value engineering research papers

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• Published: 02 June 2022

Evaluation of influential value engineering factors on the function of interchanges: case studies in Iran

• Mohsen Jafari Nigjeh 1 &
• Nima Amani   ORCID: orcid.org/0000-0002-8551-9519 2  

Journal of Engineering and Applied Science volume  69 , Article number:  48 ( 2022 ) Cite this article

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In Iran, recently in large-scale projects, the science of value engineering has become very important and in order to show the significant impact of value engineering results in increasing efficiency and reducing the cost of projects, the reason for choosing this research. The experience of value engineering in the developed countries shows the significant effect of this method in the value optimization of plans and projects. This research, too, aims to evaluate the effects of value engineering on the performance of interchanges in Iran, so that with comparing their results, the most important qualitative parameters can be determined, and also to present a suitable model for cost reduction per each IRR of the value engineering study’s expenses. The first part of this research will give a short introduction on value engineering and its job plan, and then, 8 value engineering studies of interchanges in the cities of Tehran, Karaj, Shiraz, and Mashhad will be qualitatively and quantitatively analyzed, and their results as a qualitative and quantitative evaluation (including construction cost, total value index, and relative value index) will be compared with the base plan of the projects and also with each other. Finally, the outcome of the research will show that among the evaluation parameters, the acquisition parameter plays a significant role in choosing the best design in value engineering, some part of which depends on the employer’s view in this regard. In the next step, the best improvements were in reducing travel time and in increasing safety, and also the most neglected parameter in the value engineering analyzed here is environmental improvements, which demands especial attention. From this research, it can be concluded that per every one IRR of expenditure on performing value engineering studies, the total cost of interchange projects in Iran is reduced by 334 IRR.

Introduction

Value engineering is a systematic attempt which aims to study and analyze all of the operations of a project from its conception up to the design stage and then initiation and utilization and is considered as one of the most effective and important economical methods in the engineering field [ 24 ]. In the framework of project management, value engineering is paying attention to all of the components of the design and at the same time not taking for granted any part of the project [ 1 ]. The goal in VE is reducing the time needed to reach initiation stage without increasing the cost or lessening the quality of the work [ 9 ]. Project management is planning and directing the project in a framework of specific time, cost, and quality constraints toward its specific results [ 6 ]. Standing (2001) considers VE as a project with minimum cost which includes due assessment and elimination of unnecessary costs without having detrimental effects on safety, quality, reliability, implementation method, and delivery [ 32 ]. In this paper, 8 VE case studies on Iranian interchanges are used, enabling this research to identify and analyze the benefits of the successful application of value engineering in enhancing the quality of interchange projects. The goal of this paper is the qualitative and quantitative measurement and evaluation of the effective factors of VE projects on interchanges and extracting the shared effects in order to increase quality, performance, and value index of such projects. This research will determine the amount saved for the cost of value engineering projects in Iranian interchanges. To conduct this research, extensive studies in this field have been conducted from various sources, and their impact and results have been used (Elhegazy H (2021) [ 13 ], Elzarka et al., (2016) [ 11 ], Chakraborty et al., (2020) [ 10 ], Chakraborty et al., (2016) [ 11 ], Dell'isola (2017) [ 12 ], Elmousalami [ 18 ], Kumar and Gururaj [ 22 ], Swei et al. [ 33 ], Elhegazy et al., (2020, 2021, 2022) [ 14 , 15 , 16 , 17 ], Wang et al., (2017) [ 36 ], Zhong et al., (2022) [ 37 ]).

Value engineering basics

Value engineering is a powerful methodology when it comes to problem-solving, cost reduction, and at the same time improving performance and quality, which by identifying and enhancing value indices, and applying creativity, increases client satisfaction and the value of investment.

Value engineering job plan

A value engineering job plan is comprised of three stages (Iyer, 2009) [ 20 ]:

Value engineering or value analysis workshop

After study

Each stage having a job plan as is shown in Table 1 [ 8 ].

Value analysis plan includes operational implementation steps of value methodology [ 23 ]. In fact, that which Miles and the people coming before him were doing was the application of this key stage. The second stage has 6 steps which are applied in order as depicted below. The International Society of American Value Engineers (SAVE) has proposed the job plan depicted in Table 2 for the value engineering workshop (SAVE international, 2017 [ 31 ]; Kolano & Eta, 2015 [ 21 ];) (second stage).

The International Society of American Value Engineers (SAVE) has proposed the following procedure (Fig. 1 ) for the steps mentioned in Table 2 [ 21 , 31 ].

The schematic of the value analysis process in VE workshop

International experiences regarding value engineering interchanges

The experience of value engineering in the developed and developing countries demonstrates the significant effect that this method has on improving the value of designs and projects. International experience shows that, in a period of 8 years, 2700 value engineering studies have been carried out in the transportation sector of the USA, for which the ratio of cost reduction (with preserving or improving quality and function) to the study costs has been 113 dollars to 1 [ 9 ]. In the US road sector alone in 2008, 382 value studies were carried out. Twelve billion dollars were spent conducting these studies. The approximate total cost of these projects was roughly 30,000 billion dollars. The amount of cost reduction produced as proposals in value engineering workshops was 6500 billion dollars from which 2530 billion was approved. This reported and approved saving shows that for every 1 dollar spent on value engineering, 205 dollars have been saved [ 32 ]. The statistics given here are from the value-engineered projects funded through federal government’s budget, and if we were to include the projects value engineered by private sector contractors, the figures would have been higher. The results produced in the USA illustrate a number of pivotal points:

The strength and accuracy of the base plan (the initial design by the consultant) not only do not preclude value engineering, but it paves the way for presenting new ideas and increasing the value of the project.

Not every value engineering proposal is necessarily approved. In most cases, only 50% of them gets approval.

The reported savings only include construction costs, and if the costs accumulating in the life span of the project were included, the figures would have been significantly higher [ 2 , 32 ].

A well-designed methodological research presents the administrators and engineers of road projects, including interchanges, with the most effective approaches for solving many of the problems they encounter. Although, the growth rate of highway transportation brings more complex problems with itself, which demands special attention on the part of decision makers. All selected projects are among the major and very important projects in Iran, which have been designed and implemented by the most prominent and well-known consultants and contractors, and these projects are among the highest projects in terms of price and cost of work.

For all selected projects, the value engineering discussion has been done extensively and separately by the country’s leading expert consulting engineers, and it has not been possible to provide all the details for each project separately. Therefore, the statistics and results obtained from value engineering, which is completely accurate and specialized, have been used to conduct research, and the results obtained from the value engineering of projects have been transferred and used without any change in the analysis of relevant research data.

At first, in this research, all civil, traffic, construction, economic, value, base plan, and value engineering design studies, were gathered from their respective consulting firms for analysis. The general specifications of the projects and the consulting firms of the base plans and value are presented in Table 3 . The total cost of the base plan is comprise of the construction costs, repair and maintenance, utilization, and land acquisition [ 3 , 4 , 5 , 7 , 19 , 25 , 26 , 27 , 28 , 29 , 30 , 34 , 35 ]. In the next stage, all of the parameters will be qualitatively and quantitatively analyzed.

All selected research projects are among the large and very important and well-known projects in Iran in which value engineering is very important, and in general, it can be said that value engineering reduces costs. A total of 10 to 40% in these projects and due to the budget deficit of many construction projects in Iran, cost reduction is the most important factor in value engineering in this country.

Qualitative analysis

Considering the forms sent to all beneficiary parties of the interchange projects, the parameters deemed important by the beneficiaries and team experts were gathered, integrated, and extracted as is presented here [ 7 , 19 , 27 , 28 , 29 , 30 , 34 , 35 ].

Based on the project value studies of the projects under analysis done by the consulting firms (named in Table 3 ), the evaluated qualitative parameters of the projects are presented in Table 4 .

The value engineering team in each project, noting the location, type, and the particular conditions of that project, compares the qualitative parameters using binary AHP method. Then, for each parameter, a numerical score is calculated, and finally, the parameters having the highest score are chosen for analysis and proceeding with the value engineering process. The scoring is done by binary comparison of the parameters and giving each one a score ranging from 0 to 10, where the sum of score for both of the parameters is 10. The priority and importance of the parameter determine the score it receives, that is, the higher its priority and importance, the higher the score it receives. In such manner, using the binary method, all of the parameters are separately compared, their individual score is calculated, and finally those having a score higher than 5 are chosen. After comparing the parameters using AHP, based on the score they received by the value engineering team of each project being studied, the chosen qualitative parameters were analyzed. Then, all the qualitative parameters of each project were analyzed (according to the Table 4 ), and those parameters which were shared by at least 50% (i.e., 4) of these projects and were among the important qualitative factors were extracted. In the following Table 5 , points (rates) of qualitative parameters in each project can be seen.

In the next stage, we calculated the average score of each parameter for these projects. Table 6 shows the average for the influential parameters determined in the previous stage.

Keeping in mind that the base plan score is evaluated as 5, therefore, by calculating the ratio of the base plan score to that of the calculated average scores from Table 6 , the optimization rates of the qualitative parameters, as shown in Table 7 , are obtained.

Therefore, it can be concluded that the priority of qualitative parameters of interchanges in Iran (according to the conducted case study) and their optimization rate is as shown in Table 8 .

Based on the conducted research, it can be concluded that in Iran, when constructing interchanges, among the qualitative parameters, the acquisition facility has an especial importance for the employer (in most cases the government), as it has the highest optimization rate among the qualitative parameters leaving them behind with a significant margin. It is worth mentioning that in analyzing each project separately, acquisition facility had the highest optimization rate compared to other parameters. This goes to demonstrate that in many of the value-engineered projects, considering the high cost of land and buildings in Iran, the employers in choosing the best option among the value-engineered options, put forward by the value consulting firm, go for the one with no or the fewest number of acquisitions or ultimately the one having the highest acquisition facility.

The other point worthy of attention in this research is that reducing environmental degradation occupies the last place (i.e., it has the least priority) among the important and influential qualitative parameters with an optimization rate of zero. Unlike other countries which place a significant importance on preserving and improving the environment, unfortunately, in Iran, this issue is neglected especially by the governmental employers, which requires reconsideration and more attention.

Quantitative analysis

All of the information and documentations related to the base plan and value engineering studies of the aforementioned projects were analyzed and studied, and the data including the construction cost of the base plan, the construction cost of the top value engineering plan, the total cost of the base plan (comprising of the construction costs, repair and maintenance, utilization, and land acquisition), and the total cost of the value plan of the projects were extracted and are represented here in Table 9 .

For each project, the construction cost reduction is calculated by finding the difference between value plan’s construction cost and construction cost from base plan. Also, the total cost reduction is the result of subtracting the total cost of value plan from the total cost of the base plan (in Table 10 ).

Afterwards, using the value engineering contracts of each project, the cost of value engineering was extracted, and the total sum of the cost of value engineering studies was obtained, and next, the total sum of cost reduction was calculated. To calculate the profit gained for every IRR spent on value engineering (saved amount), the sum of total cost reduction must be divided by the sum of value engineering costs. The resulting figure represents the amount saved for every IRR of value engineering cost (in Table 11 ).

As it can be observed in the table above (Table 9 ), in value engineering interchanges in Iran for every IRR of the cost of value engineering studies, 586 IRR is saved from the total cost. But in order to improve the accuracy of the research, taking into consideration that the initial construction cost of these interchange projects are not identical, and also the size of the projects, a range must be chosen so that the figures obtained can be realistic and credible. To do so, a variance and a standard deviation were calculated using formula 1 , so that the intended range could be calculated using formula 2 . Then eliminating the data outside of that range, the figure given above was fine-tuned.

X m : Total average of saving for every IRR of the cost of value engineering studies of the projects

X i : The amount saved for every IRR of the cost of value engineering study of each project

n : the number of projects

Obtaining the range of the research, it is determined that one of the projects is located outside of the domain, therefore, leaving the numbers for the said project out of the calculations, the amount saved is recalculated as follows (formula 3 ).

The total saving of the projects located inside the obtained range is 2340 IRR, which if it is divided by the number of projects located inside the range, on average, the saving would be 334 IRR for each project. Consequently, it is revealed that for every IRR cost of the value engineering studies conducted for these interchange projects, 334 IRR is saved from the total cost of interchange project in Iran. Accordingly, it can be deduced that for the interchange project in Iran, on average, for every IRR spent on value engineering, 334 is saved from the total cost of these kind of projects, which testifies as to the necessity of value engineering and its tangible effects on interchange projects in Iran. The international experience shows that in federal highway projects, the amount saved for every dollar of the cost of value studies in the year 2009 has been 99 dollars; 2010, 146 dollars; 2011, 80 dollars; 2012, 96 dollars; and in the year 2013, 118 [ 32 ]. In the US road sector alone in 2008, 382 value studies were carried out. Twelve billion dollars was spent conducting these studies. The approximate total cost of these projects was roughly 30,000 billion dollars. The amount of cost reduction produced as proposals in value engineering workshops was 6500 billion dollars, from which 2530 billion was approved. This reported and approved saving shows that for every 1 dollar spent on value engineering, 205 dollars have been saved [ 32 ].

Conclusions

The outcome of this research demonstrates that among the evaluation parameters, the acquisition parameter in interchange projects plays a significant role in choosing the top value engineering plan. As it was mentioned, this partially depends on the views that the employers hold. The highest improvements accomplished were in reducing travel time and increasing safety. The most neglected parameter in the value engineering plans evaluated was improving the environment which demands especial attention. Also, from this research, it can be concluded that for every IRR spent on conducting value engineering studies, 334 IRRs are saved from the total cost of interchange projects in Iran.

The value engineering study should be started in the design stage of interchange projects.

Availability of data and materials

The datasets used and/or analyzed during the current paper are available from the corresponding author on reasonable request.

Abbreviations

The analytic hierarchy process

United States dollar

• Value engineering

Iranian rial

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Construction Management, Department of Civil Engineering, Chalous Branch, Islamic Azad University, Chalous, Mazandaran Province, Iran

Mohsen Jafari Nigjeh

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MJ carried out the research methodology, developed the computational technique and performed the investigation and validation of the analytical work, and wrote the manuscript draft. NA performed the conceptualization of the research idea, participated in the interpretation of the results, reviewed the edited manuscript, and supervised the whole project. Also, he contributed to the discussion of the results and visualization of the final manuscript. The author(s) read and approved the final manuscript.

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Nigjeh, M.J., Amani, N. Evaluation of influential value engineering factors on the function of interchanges: case studies in Iran. J. Eng. Appl. Sci. 69 , 48 (2022). https://doi.org/10.1186/s44147-022-00100-9

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• Interchanges
• Cost reduction
• Improving qualitative parameters

State of Art in Value Engineering: A bibliometric based review for future research

Manufacturing Technology and Research (An International Journal)

9 Pages Posted: 18 May 2021

Anbesh Jamwal

Date Written: May 16, 2021

In today’s world the market is very competitive and complex to survive innovative identity and values propositions of the company. In several past decades the evolution of value engineering as a successful and important tool to company is remarkable and still there is an opportunity which is needed to be addressed. This review intends to identify how value engineering helps in improvement of design and cost of developed products. In this review numbers of value engineering published articles are extracted and investigated from top journals, productive articles and citied conferences papers. The considered papers to summarize the research growth in value engineering have been published from year 2001 to 2020. Our analysis of this papers provides informational insight on value engineering and might be resourceful for those who are research oriented towards value engineering.

Keywords: Value engineering, Cost reduction, Value Management, Value Methodology, Value Analysis, bibliometric analysis

JEL Classification: L00

Suggested Citation: Suggested Citation

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Anbesh jamwal ( email ).

Department of Mechanical Engineering MNIT Jaipur Jaipur, UT 302017 India

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Value engineering application in a high rise building (a case study in Bali)

Putri Arumsari 1 and Ricco Tanachi 1

Published under licence by IOP Publishing Ltd IOP Conference Series: Earth and Environmental Science , Volume 195 , The 2nd International Conference on Eco Engineering Development 2018 (ICEED 2018)5–6 September 2018, Alam Sutera Tangerang, Indonesia Citation Putri Arumsari and Ricco Tanachi 2018 IOP Conf. Ser.: Earth Environ. Sci. 195 012015 DOI 10.1088/1755-1315/195/1/012015

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Finding affordable alternatives material to replace the initial design of the architecture work of a commercial high rise building project without reducing the value of the material itself. Then analysing the cost saved from applying the value engineering in the architecture work of a high rise building. Calculation through the value engineering process was conducted. A commercial high rise building in Bali was chosen as a case study to implement the value engineering process in the design stage. The value engineering stages included calculating the cost/worth function, analysing through Function Analysis System Technique (FAST), innovation and creativity stage, evaluation of the life cycle cost, decision analysis and last the decision making. The research found that by applying value engineering through several items of work in a commercial high rise building it could save up to up to 8% of the total cost of the architecture work. The architecture work that was analysed through value engineering were the wall work, the door work, the floor work and the sanitary work. These were found to be the most effecting result from the Pareto calculation. The application of value engineering is this research was only done in one commercial high rise building in Bali. The work analysed was also only the architecture work. Therefore other types of construction project might result in different value of cost saving. This research identifies the cost saved through the application of value engineering in a commercial high rise building in Bali, which emphasizes on high quality materials in the architecture work. This research will contribute to the study literature of value engineering with a case study of commercial high rise building.

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VALUE ENGINEERING: A VITAL TOOL FOR IMPROVING COST & PRODUCTIVITY

To face the new business challenges, Companies have to increasingly depend upon the implementation of better management practices. The Practices, which are adopted by them so far for achieving their position in the economy, will need to be reoriented. Business organizations have now to look for innovative ways to cut costs, improved quality and have more customers’ orientation. In this perspective, when the directorial concepts are value and satisfaction. The role of value engineering as cost reduction and control technique becomes vital for an organization. The present paper discusses in detail value analysis/value engineering, as an important tool for reducing the cost. The paper takes an overview of importance of cost reduction and the link between cost reduction and value analysis. The study further takes up the necessity of value engineering in today’s scenario and tells when to apply. In the end of the paper also explains the approach and methodology of engineering. Few examples of corporates have also been provided that have achieved cost reduction by adopting and implementation technique of VE.

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Bridge construction is a very important infrastructure because it connects two separate places due to several conditions. This research was done under the bridge of Lamnyong Bridge doubling construction in Banda Aceh, by providing the most economical costs but still meet the strengthen requirements specified or without losing the value and function of the building. The scope of this research is limited to the implementation cost of under Lamnyong Bridge construction structural works, such as: foundation, bridge abutments and pillars works. The method used in this research is value engineering method that is analysis oriented to evaluate the function of construction project planning of the work. This analysis has a systematic and focused approach in evaluating the object being surveyed. The objective of this study is to apply value engineering to the construction implementation method of concrete conventional bridge construction. There are some implementation phases in value engineering, they are information phase continuing to identify the budget starting from the highest to the lowest prices by using Pareto law distribution graph the creative phase by using cost/worth method, analysis phase and the last is recommendation phase. From the value engineering application of the early design with the budget is 72,486,508,196.71 IDR then carried out some alternatives, they are alternative I obtained the cost is 40,616,598,222.56 IDR or it is 43.97% cost saving, for alternatives II obtained the cost is 41,699,143,562.90 IDR or it is 42.47% cost saving and alternative III obtained the cost is 41,243,208,716.90 IDR or it is 43.10% cost saving. Therefore value engineering method can optimize project cost saving in order to improve effective development quality.

ABSRACT This paper gives a prospective view of QFD methodology and reach to product development. The intention is to enable the customers to initiate and interact with the product development process, which gives a competitive factor by augmenting customer value and strong relation to customers, which increase belief in the company in competitive business. The customer driven product development (CDPD) is found through an explored research design along with global industrial markets and its customer. Using the Tool QFD which translates customer needs into technical requirements, analyzes and assesses the competitor's product parameters, this paper recommends a conceptual model of QFD implementation focussing to establish quality parameters in the steel rolling industry. The customer is involved in all the phases of process development viz. Product planning, design, process planning, operations planning, and forms of a new derived system to the customer. In this paper, the product square bar of hot rolled steel plant has been studied for the ranking of customer requirements and prioritized design. Using this method keeps customer focused reduction in the product development cycle, increases customer satisfaction.

This study examined the impact of service providers’ emotional intelligence on service quality and customer satisfaction in the telecommunication sector of Ghana. A quantitative research technique was adopted to test hypotheses whose conclusions could be generalised over the telecommunication sector in Ghana. Probability sampling methods were used to select 384 each of customers and employees of all telecommunication firms in Ghana. Data were collected using a self-administered questionnaire. Data were analysed using Pearson’s correlation test, partial correlation test, multiple linear regression and Analysis of Covariance (ANCOVA). Findings of this study indicated that emotional intelligence is highly positively related to service quality (r = .889, p = .000) and customer satisfaction (r = .573, p = .000). Also, emotional intelligence moderates the relationship between service quality and customer satisfaction. Service quality and emotional intelligence make an interactive effect on customer satisfaction. Emotional intelligence and service quality significantly predict customer satisfaction (p < .05) and account for 57.3% of variance. More importantly, emotional intelligence significantly predicts service quality (p < .05) and accounts for 79% of variance. It is concluded that emotional intelligence significantly predicts both service quality and customer satisfaction, though it predicts service quality more strongly.

International Journal of Mechanical and Production Engineering Research and Development (IJMPERD)

Economic growth and environmental protection are mutually conflicting in nature. Both issues are to be addressed simultaneously in order to sustain green supply chain management. Suppliers and customers have to focus these aspects in deterring their business relationship. An effort is made here to deploy multi criteria technique fuzzy ahp, Taguchi loss function and fuzzy multi objective technique to ascertain the quantity to be ordered on the supplier. Case study is conducted. The results are impressive.

Isaac Zeb-obipi

Productivity increase by means of a workstudy in a manufacturing industry is the area of interest in this project. The project was conducted life , wherein numerous types of tools and techniques were employed to improve the efficiency and productivity of the industry. In essence, our project deals with the small-scale manufacturing industry. This concerned company in the project manufactures and supplies sheet metal components, including cyclones, electrical panel, stainless steel online humidification system and its components, and controlling panel. In addition, the company is a vendor of Nail Strip Jumbo, which is utilized in ginning machines. This live project applied work study methods to improve the practices in the industry, in addition to ascertaining and rectifying problems associated with the production process. The employment of such techniques improved production by reducing production time and processes involved, as well as an increase in the production rate. This project highlights the advantages of adopting such an efficient process.

Considering the fact of soaring improvement of Internet, e-trade sites nowadays habitually one need to labor by means of datasets logging that are up to a couple of terabytes measuring in dimension. The most effective method to expel chaotic information opportune with minimal effort and discover helpful data is an issue we need to confront. The mining procedure includes a few stages from pre-handling the crude information to building up the last replicas. To deal with the issue of extricating and keeping up countless profiles from substantial scale information, initially the researchers depict the distinctive versatile usage of the projected system. At that point the researchers would witness the difficulties they confronted in the execution. Recommender System consists of arrangement a feature which familiarizes the client inclination, faith, expectation precision, certainty and so forth. Utilizing information and students' inclinations prescribe things by means of different connections to decrease Stability issue of Recommender System. Content oriented channel has been projected to concentrate client report in light of client area attention. To discover pertinent item Similarity events and characterization has been utilized to order the item as per client question. Here Recommendation is achieved based on artificial intelligence. Artificial intelligence is creating a computer, a computer controlled robot, or intelligent software think, in the related way the intelligent humans think. We apply this technique for timely recommendation. It recommends the product and related accessories based on user earlier searching. The client could provide the remarks about the items. This study suggests the wistful examination of the client concerning the item in view of the remarks. The researchers have utilized guideline parts examination so as to gather Product nostalgic investigation points of interest, get-together remarks overall Product wistful examination with expect individuals passionate distribution with regard to Product assessment sentiments. Bunching group Product nostalgic independently on the grounds that passionate slant separate gathering for effortlessly distinguish individuals however, recommendation, emotion, as concerning Product feelings in remark. The covering apportioning bunching has been utilized to aggregate the information. The covering parcel incorporates the fuzzy and SVM. Bunching assembled to different wistful, for example, great, awful, amazing, poor, and so forth subsequently intended for tweeting division in a gathering, known as Sentiment. Through part remarks interested in important portions, the optimistic or pessimistic data has been all around safeguarded and effectively extricated through the downstream functions, it utilizing Multilayer observation strategy. Separating, Sentiment locates the ideal portion of a tweet through amplifying the entirety of the tackiness points of its competitor fragments. The tackiness point regard as the likelihood of a section being an expression similar to and the likelihood of a fragment being an expression inside the gathering of not at all like. At long last the outcome would be viewed as Product worth has been in terms of how a great deal similar to or dissimilar to individuals wistful practices and feeling have been precision with excellence.

José G. Vargas-Hernández

The purpose of the study is to identify factors which are evaluated formally for the analysis of economic units (EUs) as the basis for determining strategies for its development. During the study the model is used as an official to diagnose the current state of the reference units is defined. The investigation includes a comparison of some national and international studies, which show the similarities and differences on the subject treated. This research has identified that the

As the business environment is becoming more competitive and unstable than ever before, organizations are seeking to gain competitive advantage through enhanced employee performance. To achieve this, organizations have recently focused on the human resources practices through more innovative approaches. The aim of the present study was to establish the effect of human resource management practices on employee performance in large scale tea farms in Kenya. The study specifically looked at the effect of selected practices and how they impact on the overall employee performance. The study employed a descriptive research design specifically a case study of tea firms. The target population was employees of all three large scale tea farms in Kuresoi South District. The target population was 2750 employees and the study used cluster sampling due to the geographical coverage of the tea farms to have a representative sample of 96 employees. The study used both qualitative and quantitative data approach in its data collection. The collected data was summarized and analyzed using both descriptive both descriptive and inferential statistics and then presented in tables, graphs and charts. The findings indicated that there was a strong relationship between employee training, employee welfare practices, compensation and job security measures and employee performance. Compensation and employee welfare practices with correlation coefficients of r = 0.673 and r = 0.485 were found to have the most significant effect on employee performance. The study recommended that tea farms need to put in place effective compensation schemes and progressive employee welfare practices in order to enhance employee performance. Further, the large scale farms need to enhance both their employee training processes and job security measures in a bid to motivate their employees if employee performance is to be increased.

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In Situ Monitoring and Defect Diagnosis Method Based on Synchronous Compression Short-Time Fourier Transform and K-Singular Value Decomposition for Al-Carbon Fiber-Reinforced Thermoplastic Friction Stir Lap Welding

• Original Research Article
• Published: 15 August 2024

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• Yibo Sun 1 ,
• Haiwei Long 1 ,
• Siyu Zhao 1 ,
• Yuan Zhang 1 ,
• Jianning Zhu 2 ,
• Xinhua Yang 2 &
• Libin Fu 2  

Lightweight is one of the key technologies and research hotspots in the fields of aerospace and railway, which makes the demand for the joining of dissimilar materials such as Al-based and carbon fiber-based (Al-CFRTP) materials. In friction stir lap welding (FSLW), weld defects are easily generated due to differences in physical and chemical properties. In this paper, an in situ monitoring and defect diagnosis method for FSLW of Al-CFRTP is proposed. In this method, time–frequency features are extracted by synchronous compression short-time Fourier transform (SSTFT). The welding status is classified by the K-singular value decomposition (KSVD), including two welding states and three welding defects. From the SSTFT results, the frequency band shifts from 23 to 10 kHz accompanied by the nugget collapse defects and shifts from 25 to 17 kHz accompanied by the flash and surface galling defects. From the KSVD results, the recognition accuracy of defects reaches 90% based on KSVD prediction model. The computation speed is 100 times faster than neural networks with nearly same recognition precision. The superiority of this method is the precise characterization description of FSLW defects from time–frequency domain features based on SSTFT and the fast processing speed in the classification by KSVD.

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Acknowledgments

This work was supported by the National Science Foundation of China (52005071, 51875072), Natural Science Foundation of Liaoning Province (2022-MS-342), and Liaoning Provincial Department of Education General Project (LJKZ0478).

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Yibo Sun, Haiwei Long, Siyu Zhao & Yuan Zhang

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YS involved in conceptualization, writing original draft; HL took part in investigation, methodology, programming; SZ participated in manuscript review; YZ involved in experimental data curation and manuscript review; JZ took part in supervision, resources; XY carried out project administration. All authors have read and agreed to the published version of the manuscript.

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Sun, Y., Long, H., Zhao, S. et al. In Situ Monitoring and Defect Diagnosis Method Based on Synchronous Compression Short-Time Fourier Transform and K-Singular Value Decomposition for Al-Carbon Fiber-Reinforced Thermoplastic Friction Stir Lap Welding. J. of Materi Eng and Perform (2024). https://doi.org/10.1007/s11665-024-09930-5

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Exploring Agrobacterium -mediated genetic transformation methods and its applications in Lilium

• Xinyue Fan 1 &
• Hongmei Sun 1 , 2  

Plant Methods volume  20 , Article number:  120 ( 2024 ) Cite this article

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As a typical bulb flower, lily is widely cultivated worldwide because of its high ornamental, medicinal and edible value. Although breeding efforts evolved over the last 10000 years, there are still many problems in the face of increasing consumer demand. The approach of biotechnological methods would help to solve this problem and incorporate traits impossible by conventional breeding. Target traits are dormancy, development, color, floral fragrance and resistances against various biotic and abiotic stresses, so as to improve the quality of bulbs and cut flowers in planting, cultivation, postharvest, plant protection and marketing. Genetic transformation technology is an important method for varietal improvement and has become the foundation and core of plant functional genomics research, greatly assisting various plant improvement programs. However, achieving stable and efficient genetic transformation of lily has been difficult worldwide. Many gene function verification studies depend on the use of model plants, which greatly limits the pace of directed breeding and germplasm improvement in lily. Although significant progress has been made in the development and optimization of genetic transformation systems, shortcomings remain. Agrobacterium -mediated genetic transformation has been widely used in lily. However, severe genotypic dependence is the main bottleneck limiting the genetic transformation of lily. This review will summarizes the research progress in the genetic transformation of lily over the past 30 years to generate the material including a section how genome engineering using stable genetic transformation system, and give an overview about recent and future applications of lily transformation. The information provided in this paper includes ideas for optimizing and improving the efficiency of existing genetic transformation methods and for innovation, provides technical support for mining and identifying regulatory genes for key traits, and lays a foundation for genetic improvement and innovative germplasm development in lily.

Flowers are not only a globally important agricultural industry with great economic benefits but also necessary agents for mental health in people's daily lives. Many countries have given intensive attention to the development of the flower industry. At present, the world's flower cultivation area is 22.3 hm 2 , and the international trade volume of flowers is expanding (AIPH, https://www.floraldaily.com ). Lily is a typical perennial herbaceous bulb plant with more than 100 wild species and more than 9,000 varieties worldwide; additionally, this plant has high ornamental value, and the share of lily as a cut flower in the global flower market is increasing annually [ 1 ]. Some lily varieties are edible and have high medicinal value, and their extracts are rich in antioxidant and anti-inflammatory components, resulting in widespread use in medicine, functional food and cosmetics [ 2 ]. It is therefore unsurprising that lilies are the focus of much bulbous flower research. Improving the quality of seed balls and cut flowers has always been a key goal in the global lily industry [ 3 , 4 ]. The target traits are dormancy, development, colour, floral fragrance and resistance to various biological and abiotic stresses to improve the quality of bulbs and cut flowers in planting, cultivation, postharvest, plant protection and marketing. Crossbreeding can quickly fuse good traits and easily produce heterosis. However, lily has a complex genetic background, high heterozygosity, and it is extremely difficult to carry out genome processing, because it is one of the plants with the largest genome, with nearly 30 Gb of genetic information. The long cycle of cross-breeding requires a lot of manpower and material resources. In many cases, sexual incompatibility is also an obstacle in the crossbreeding of lily. Likewise, physical and chemical mutagenesis methods are also highly uncertain. With the rapid development of molecular biology technology, genetic engineering has received increasing amounts of attention. Using molecular methods to improve germplasms can not only lead to the creation of new traits but also increase the efficiency and accuracy of breeding [ 5 , 6 , 7 ] (Fig.  1 ).

Strategies for developing new varieties of lily using different plant breeding tools. A Physical mutagenesis (in which mutants are created by exposing seeds or bulblets to radiation). B Chemical mutagenesis (treatment of different explants with chemical agents to obtain mutants, such as EMS mutagenesis). C Traditional crossbreeding has led to the cultivation of new lily varieties. D Transgenic breeding

Genetic transformation is an important part of genetic engineering technology, and the main goals of flower genetic transformation are as follows: (1) genetic transformation for basic research on a single gene, gene family or gene regulatory network and (2) the application of basic research results as the theoretical basis for improving flower traits and creating new varieties. Plant genetic transformation includes target gene selection, delivery, integration into plant cells, and expression and, ultimately, the production of a complete plant after numerous processes [ 8 ]. Although genetically modified plants were obtained in 1983, the genetic transformation of bulb flowers such as lily has long been considered difficult or impossible [ 9 ]. With progress in the field of lily research, an increasing number of genes, including genes related to major factors involved in regulating various life activities, responding to various biological and abiotic stresses, and responding to various environmental signals, have been identified. However, many gene function studies still depend on the heterologous transformation of model plants such as Arabidopsis and Nicotiana benthamiana (Fig. S1) . In 1992, Cohen conducted the first genetic transformation experiment in lily through Agrobacterium -mediated transformation and detected foreign genes in the calli [ 10 ]. However, the low efficiency, difficulty of regeneration, and difficulty of integration into the lily genome remain obstacles to overcome. With increasing basic research on lily plants, instantaneous transformation based on virus induction has gradually become the first choice of many researchers because this method is simple and fast, and the research cycle is only a few hours or days. However, because these methods cannot enable integration into the genome and are sometimes limited to a single tissue, it is difficult to provide sufficient evidence for gene function, and the research results are uninformative and unfavourable for further application in breeding work. Agrobacterium -mediated transformation, particle bombardment, PEG and electric shock are common methods of plant genetic transformation at present (Fig.  2 ). Compared with other plant transgenic methods, Agrobacterium -mediated plant genetic transformation remains the most common and widespread lily transgenic strategy because of its advantages of high transformation efficiency, few transgenic copies, and stable transfer of integrated genes into offspring after continuous optimization and updating [ 11 ].

Common methods for the genetic transformation of lily. A Agrobacterium -mediated stable genetic transformation B Virus-induced transient gene silencing (VIGS). C Particle bombardment. D Pollen magnetic effect method

In the past 30 years, many researchers have attempted to improve and create technology by adjusting or changing various parameters and operating methods and have accumulated considerable valuable experience (Fig.  3 ). Agrobacterium is a gram-negative bacterial genus that is widely distributed in soil. At the beginning of the twentieth century, the principle that natural pathogens can infect plants through wounds was gradually elucidated. The main mechanism is the delivery of tumorigenic DNA molecules (transfer DNA or T-DNA) into plant cells through wounds in infected plants; these molecules are eventually integrated into the host genome and stably transmitted to the next generation of the plant through meiosis [ 12 , 13 ]. The ability of Agrobacterium to integrate its own DNA into the host genome is primarily determined by the Ti plasmid [ 14 ], which can be modified by the insertion of target genes into the T-DNA region. With the help of the transferability of this region, genes can be introduced into plants by Agrobacterium infection and incorporated into plant genome, after which transgenic plants can be generated by cell and tissue culture technology [ 13 ]. Like most plant genetic transformation methods mediated by Agrobacterium, the genetic transformation procedures for lily mainly include vector construction, selection and culture of explants, preculture, Agrobacterium infection, coculture, resistance selection and transgenic plant regeneration (Fig.  2 ). At present, in addition to calli induced by roots, petals and leaves, scales and embryonic calli are common explants used for genetic transformation in lily [ 15 , 16 , 17 ]. With the continuous updating and optimization of genetic transformation technology for lily, functional verification of several genes by heterologous transformation of model plants has gradually increased, and many genes that regulate desirable traits in lily have been identified [ 3 , 17 , 18 , 19 , 20 ]. Nevertheless, stable and efficient transformation of target genes has been achieved in only a few lily varieties, and the success rate of genetic transformation of some lily varieties is still low.

Timeline of several major discoveries, applications and breakthroughs in the history of lily genetic transformation

In this paper, the development of genetic transformation technology for lily plants over the past 30 years is reviewed, the factors and key technical points restricting the efficiency of genetic transformation in lily are described, the problems and limitations associated with the genetic transformation of lily are summarized, and the prospects for application and improvement are discussed. The purpose of this paper is to provide a technical reference for establishing a stable and efficient genetic transformation system for lily and to lay a foundation for directional breeding and genetic improvement of key characteristics.

Factors affecting the genetic transformation of lily

Many factors affect lily regeneration and transformation. Genotyping is one of the key problems affecting the success of transformation [ 16 ]. Although genetic transformation systems have been established for different lily varieties, major differences exist between different genotypes [ 21 ]. Under the same conditions during the genetic transformation process, the genotype determines the difficulty of using Agrobacterium to successfully infect lily. At present, stable and efficient genetic transformation has been successfully achieved for few lily varieties (Table  1 ). Since the use of Agrobacterium -mediated genetic transformation of lily has been reported, several studies have aimed to optimize the transformation system or establish methods suitable for different lily species, including Lilium formolongi [ 22 , 23 ], Lilium longiflorum [ 24 , 25 , 26 ], Lilium pumilum DC.Fisch. [ 26 ] and the Oriental hybrid Lily [ 15 , 16 , 27 , 28 ]. Due to the strong genotypic dependence and difficult regeneration of explant materials after transformation, most related research results are restricted to certain genotypes [ 16 , 25 , 27 , 28 ]. Yan et al. [ 26 ] established a stable and efficient transformation system through somatic embryogenesis and adventitious bud regeneration in Lilium pumilum DC. Fisch. and Lilium longiflorum . After method optimization, the transformation efficiency reached 29.17% and 4%, respectively. Although the transformation efficiency in 'White Heaven' is still low, it is relatively stable and can be regenerated within 1 month. Song et al. [ 17 ] improved the original transformation system by adjusting the pH and CaCl 2 concentration of the medium; the number of resistant plants increased by 2.7–6.4 times, the number of positive lines increased by 3–6 times transformation, and the genetic transformation efficiency increased by 5.7–13.0%. In the latest study, the genetic transformation system of Oriental hybrid lily was further optimized, and the efficiency was increased to 60% by screening for the lethal concentration of antibiotics, the concentration of the bacterial solution and the duration of infection.

Good explant material is the basis of plant genetic transformation. The success of transformation depends on the selection and totipotency of explants [ 32 ]. Researchers have tested different explants for genetic transformation of target genes based on the Agrobacterium system (Table  1 ). Calli generated from floral organs, scales, leaves, or seeds have been used for genetic transformation in most lily hybrids [ 27 , 33 , 34 ]. Related studies have shown that filamentous calli have a faster growth rate and may be more susceptible to Agrobacterium infection [ 35 ]. In the Oriental hybrid lily ( Lilium cv. Acapulco), an Agrobacterium -mediated lily transformation system was successfully established by using filament-induced filiform calli as explants. Although transient expression of the GUS reporter gene could be detected by root–, leaf–, stalk–, ovary– and anther-induced callus infection, no positive transgenic tissues or plants were obtained [ 27 ]. In recent years, several other explants have been developed and offer additional possibilities for improving transformation efficiency. Liu et al. [ 24 ] discussed the effect of the direct regeneration pathway and the callus regeneration pathway on the transformation efficiency in Agrobacterium -based genetic transformation experiments using stem segments induced by lily scales as explants. Notably, when stem segments were used as explants, adventitious buds obtained via the direct regeneration pathway after coculture significantly increased the regeneration rate of resistant plants and decreased the gene escape rate [ 24 ]. Another study showed that the direct use of scales as transformation explants did not significantly improve the transformation rate but did greatly shorten the genetic transformation cycle [ 26 ]. Different explant materials have their own advantages. Cohen and Meredith [ 10 ] used a particle bombardment approach to carry out lily genetic transformation and reported that the ability of embryonic calli to accept foreign genes was 50–70 times greater than that of ordinary calli. Embryogenic calli are composed of many embryogenic cells, and each cell has the potential to develop into adult somatic embryos. Therefore, using embryogenic calli as explant materials can result in a more stable transformation population with a lower chimaerism rate, which is very important for the research and development of plant genetic transformation [ 36 , 37 , 38 , 39 ]. Mercuri et al. [ 25 ] induced embryonic calli using the styles and peduncles of Lilium longiflorum ‘Snow Queen’, and they were found to be highly competent for transformation. Recently, two studies reported an efficient protocol with high transformation efficiency for Lilium pumilum DC.Fisch. using embryonic calli. Despite their long transformation cycle, embryogenic calli are the most common explant material for the genetic transformation of lily due to their high cell proliferation rate and genetic stability [ 17 , 26 ].

Strains of agrobacterium

To date, many successful cases of stable genetic transformation of plants mediated by Agrobacterium have been reported [ 40 , 41 , 42 , 43 , 44 ]. The strain of Agrobacterium can also considerably influence the transformation frequency. With the continuous updating and optimization of plant genetic transformation technology, several researchers have attempted to improve the efficiency of plant transformation by changing various parameters, including Agrobacterium strains [ 45 ] (Table  1 ). Different plants have different preferences for the routinely used Agrobacterium strains EHA105, EHA101, LBA4404, GV3101, AGL1 and C58 [ 8 , 27 , 29 , 46 ]. In alfalfa [ 47 ], tomato [ 48 ], grasspea [ 49 ], and pigeon pea [ 50 ], LB4404 and LBA4404 were found to be more virulent and highly effective, offering higher transformation efficiency. However, the LBA4404 strain has been less frequently reported in lilies. Mercuri et al. [ 25 ] reported that LBA4404 effectively promoted the infection of embryogenic calli from Lilium longiflorum 'Snow Queen'. According to another study of genetic transformation in lily, the use of the EHA105 strain to infuse embryonic calli seemed to be more beneficial for improving transformation efficiency [ 26 ]. In recent years, the strains EHA101 and EHA105 have been more widely used in lily transformation and are considered to result in greater transformation frequency [ 16 , 18 ].

Selection of markers and reporter genes

The selection of marker genes also determines the efficiency of plant genetic transformation. They are usually delivered along with the target gene, conferring resistance to toxic compounds in plants and facilitating the growth of transformed cells in the presence of such unfavourable conditions. Normally, the marker gene and the target gene are connected to the same plasmid and delivered to the plant somatic cells via the Agrobacterium -mediated method. Suitable marker genes can help to quickly and efficiently screen many transformed materials and remove untransformed cells [ 8 ]. Conventional marker genes include the hpt gene, which encodes hygromycin phosphotransferase and confers resistance to hygromycin; the npt-II gene, which encodes neomycin phosphotransferase II and confers resistance to kanamycin, neomycin and geneticin; and the bar gene, which encodes phosphinothricin acetyltransferase and confers resistance to the herbicide phosphinothricin [ 23 , 51 , 52 ]. The type of resistance marker gene is determined in the vector, and hpt and npt-II are commonly used as marker genes for lily transformation [ 3 , 17 , 18 , 24 ]. Linking the GUS gene to a transformation vector for double or even triple marker gene screening combined with GUS histochemical staining is also an effective strategy for reducing the false positive rate of resistant plants [ 16 , 22 , 26 ] (Table  1 ).

Key parameters in the genetic transformation program

Ph of the medium.

The expression of the Agrobacterium virulence gene vir is the basis for the transformation of plant cells and the key to ensuring infection efficiency, which is strongly dependent on the pH of the medium [ 17 , 22 , 53 ]. Previous studies have shown that an acidic pH is more conducive to the expression of vir , and as the pH of the preculture or coculture medium decreases, the expression of vir is significantly upregulated [ 54 , 55 ]. Maintaining the pH at 5.2 effectively increased the genetic transformation efficiency of tomato cotyledons [ 56 ]. The virA and virG genes are switches that activate the expression of the vir gene [ 57 ]. Agrobacterium has a chemotactic system different from that of E. coli and is attracted to chemical inducers such as carbohydrates, amino acids and phenolic compounds [ 58 ]. High concentrations of chemical inducers bind to virA to induce the expression of the vir gene and trigger T-DNA transfer [ 59 ]. Acetylsyringone (AS) is a common class of natural phenolic compounds that can promote the direct entry of microorganisms into plant cells through wounds and achieve T-DNA transfer by activating the expression of the vir gene [ 58 , 60 , 61 , 62 ]. AS has been widely used in the genetic transformation of lily [ 23 , 27 ]. Although pH 7.0 is the most suitable environment for the growth of Agrobacterium , vir is more easily expressed under acidic conditions after the addition of AS, while vir expression is hardly induced under neutral pH conditions [ 54 , 63 ]. In a Lilium pumilum DC.Fisch. genetic transformation experiment, a stable pH of 5.8 in suspension and coculture media resulted in a somatic embryo transformation efficiency of 29.17% [ 26 ]. When the pH was adjusted to 5.0, the number of resistant calli increased significantly, and the transformation efficiency increased by 5.7–13% [ 17 ]. Ogaki et al. [ 23 ] reported that exogenous MES could effectively control the pH of the medium. This study further investigated the effect of adding different concentrations of MES (0, 10, 20, 50 and 100 mM) on the transformation efficiency of lily. The results showed that transient expression of the GUS gene could be observed only in coculture medium containing MES, and larger numbers of transgenic calli could be obtained by the addition of 10 mM MES buffer [ 30 ]. The above conclusions indicate that maintaining pH in the range of 5–6 values according to different varieties in the preculture and coculture stages is important for improving the efficiency of genetic transformation (Table  2 ).

Culture medium supplements

The composition of the medium is another rate-limiting factor affecting the genetic transformation efficiency of lily, and the process involves the preculture, coculture and regeneration of resistant plants. Many studies have shown that the addition or removal of certain compounds can significantly improve the efficiency of lily transformation (Table  2 ). MS medium, which contains 20.6 mM NH 4 NO 3 , is widely used in the tissue culture and genetic transformation of lily. However, the presence of NH 4 NO 3 limits the efficiency of genetic transformation in lily [ 23 , 26 , 64 ]. Previous studies have shown that virG transcription can be activated by low concentrations of phosphate [ 53 , 58 ]. When a low concentration of KH 2 PO 4 was used as a salt source instead of NH 4 NO 3 , there was no significant change in the number of regenerated resistant calli. In contrast, the complete removal of KH 2 PO 4 had a positive effect on lily transformation [ 22 ]. In a transformation study of the lily ‘Sorbonne’, it was found that the removal of KH 2 PO 4 , NH 4 NO 3 , KNO 3 or macroelements in the medium could significantly improve the transformation efficiency [ 28 ]. Montoro et al. [ 65 ] reported that Ga 2+ -free media significantly increased GUS activity in Brazilian rubber trees. However, another study concluded that the effect of GaCl 2 on plant transformation efficiency appears to be strongly dependent on genotype. Ga 2+ is considered one of the key factors involved in improving the efficiency of genetic transformation in improved lily genetic transformation systems. Increasing the GaCl 2 concentration from 0.44 g/L to 1.32 g/L significantly increased the germination coefficient of Lilium -resistant somatic embryos [ 17 ]. AS is an indispensable compound in the genetic transformation of lily, and its concentration is also a key factor; an AS concentration that is too high adversely affects T-DNA transfer [ 17 ]. Notably, researchers have found other compounds that can replace AS, and they can provide higher transformation efficiency. Chloroxynil (CX) is a class of phenolic compounds with a mode of action similar to that of AS that also improves the efficiency of treatment by activating the expression of vir. In the genetic transformation of lotus seeds, the transformation efficiency of explants treated with CX was 6 times greater than that of explants treated with AS [ 66 ]. Wei et al. [ 28 ] further confirmed the effect of CX in lily. When 4 μM CX was used, the transformation efficiency reached 11.1%, while 100 μM AS achieved only 6.6%, indicating that CX can replace AS in lily genetic transformation. Early studies showed that the promoting effect of carbohydrate substances other than glucose and xylose on vir gene activity was consistent with that of AS [ 67 , 68 ]. Further studies by Azadi et al. [ 22 ] showed that MS media supplemented with monosaccharides significantly inhibited the expression of the GUS gene, and no hygromycin-resistant lily calli were obtained. In contrast, adding sucrose significantly improved the efficiency of genetic transformation. In summary, for most series lilies, removing NH 4 NO 3 and adding an appropriate amount of AS has a positive effect on improving the genetic transformation efficiency. CX may be an excellent compound to replace AS, and it is worth further attempts in the future.

Bacterial concentration and infection time

The Agrobacterium concentration and infection time play pivotal roles in the transformation of lily. A low bacterial concentration and short infection duration will result in the failure of Agrobacterium to fully adhere to explant tissues, resulting in the inability to achieve effective transformation [ 69 ]. However, a high concentration of bacteria or long infection duration may also lead to rapid bacterial growth, which can cause severe damage to the recipient material [ 17 ]. Cell resistance varies greatly among different plant explants, and plant tolerance to different agrobacterium concentrations also differs. When the OD 600 was 0.8, the highest GUS expression rate was detected in embryogenic calli, but the percentage of resistant calli significantly decreased compared with that when the OD 600 was 0.6. For scales, the GUS expression rate and adventitious bud regeneration rate peaked when the OD 600 was 0.6. Infection time is also the key to determining transformation efficiency, and research shows that for embryonic calli and scales of Lilium pumilum, DC. Fisch, 15 min is the optimal time for Agrobacterium infection [ 26 ]. However, for embryogenic calli of the Oriental hybrid lily ‘Siberia’, an OD 600 of 0.4 was more beneficial for improving the transformation efficiency [ 18 ]. In addition, it has been reported that in the coculture process, the proliferation of Agrobacterium on the surface and around the callus increases with the removal of some elements, indicating that these elements have an inhibitory effect on Agrobacterium . Negative effects of bacterial overgrowth were observed when 10 mM MES was added to coculture media of sensitive varieties such as ‘Red Ruby’ and ‘Casa Blanca’. Therefore, screening different varieties of MES can effectively reduce bacterial growth and improve the transformation efficiency of lily [ 22 ].

Antibiotic selection

In the process of plant genetic transformation, an appropriate concentration of antibiotics can effectively inhibit the growth of non-transformed Cefatothin, and this is also a crucial step in determining the success of genetic transformation. Kanamycin, hygromycin and glyphosate have been used extensively for lily transformation due to their high availability and low toxicity, despite the occurrence of false positives in the screening of resistant plants [ 70 , 71 ] (Table  2 ). Lily explants of different genotypes have different antibiotic concentration requirements. Even within the same variety, different explant types have great differences in antibiotic tolerance [ 18 ]. Studies have shown that embryonic calli of Lilium pumilum DC. Fisch. The plants almost stopped growing and died after treatment with hygromycin supplemented at 40 mg·L −1 , resulting in extremely low growth and induction rates. However, a few scales of ‘White Heaven’ still formed complete buds under these conditions. Adjusting the concentration of hygromycin to 30 mg·L −1 reduced the browning rate of embryogenic calli by approximately 20% and significantly increased the growth and transformation rate. Therefore, 30 mg·L −1 and 40 mg·L −1 were the best hygromycin concentrations suitable for embryogenic calli and scales of Lilium pumilum DC. Fisch., respectively [ 26 ]. Another necessary antibiotic is a bacteriostatic antibiotic, which is mainly used to prevent the transformation of material from dying or difficult regeneration due to excessive Agrobacterium contamination. Cef is a common bacteriostatic agent used in plant transformation that has extensive resistance and inhibits the growth of Agrobacterium [ 72 , 73 ]. However, high concentrations of Cef can inhibit the growth of explant cells. Based on the results of studies on different lily varieties and explants, we believe that 300–400 mg·L −1 Cef may have a broad-spectrum effect [ 18 , 26 ]. The concentration of Agrobacterium may be a prerequisite for screening Cef concentrations. Even 400 mg·L −1 Cef had no bacteriostatic effect when the concentration of the bacterial solution was too high. When the OD600 of the bacterial solution is maintained within 0.2–0.4, 300 mg·L −1 Cef can have good antibacterial efficacy [ 18 ]. Therefore, it is necessary to combine the concentration of the bacterial solution with the concentration of the bacteriostatic agent during screening.

Preculture, infection and coculture procedures

Preculture, infection and coculture are the key steps in determining the success of plant genetic transformation. Previous studies have generally been conducted under the belief that the preculture of explants before transformation can effectively promote cell division so that they can maintain the best life state during infection and integrate foreign genes more easily [ 74 , 75 ]. The timing of preculture depends on the type and quality of the explants. Yan et al. [ 26 ] discussed the influence of preculture time on the transformation efficiency of L. pumilum and ‘White Heaven’. The results showed that the expression rate of GUS was lower in uncultured calli or scale explants. Similarly, compared with those of the control group, the proliferation and survival rates of the explant-treated group were significantly lower. For embryogenic calli, GUS expression and the proliferation rate were the highest in resistant calli after 10 days of preculture (66.67% and 63.33%, respectively). After 4 days of preculture, GUS expression and bud resistance began to decrease for the traumatized scales. Although the percentage of resistant buds was the highest after 2 days of preculture, a higher GUS expression rate appeared after 3 days (Table  2 ).

In lily, wounded explant materials are often more conducive to the transfer and integration of T-DNA, which can greatly improve the efficiency of genetic transformation [ 27 , 76 ]. Wei et al. [ 28 ] further confirmed this view. According to the results of Agrobacterium -mediated ‘Sobone’ genetic transformation, ultrasound treatment for 20 s can produce thousands of microwounds in explants, promote the penetration of Agrobacterium into the internal tissues of plants, and effectively improve the efficiency of transformation. The combination of heat shock and ultrasound had no significant effect. Coculture is an essential stage during which T-DNA is transferred into plant cells [ 77 ]. Therefore, coculture time is also an external factor that has been widely examined [ 75 ]. The time required for Agrobacterium -mediated gene transfer and integration into the plant genome varies widely depending on the genotype and explant type, usually ranging from a few hours to a few days [ 78 , 79 , 80 , 81 ]. Wu et al. [ 82 ] compared the transformation efficiency of bulb sections of Gladiolus under coculture for 3 days and 12 days, and the results showed that the transformation rate of coculture for 12 days was more than twice that for 3 days, indicating that a longer coculture time may benefit Agrobacterium infection and transformation. However, in Lilium pumilum DC. After more than 5 days of coculture, Fisch, which is also a typical bulbous flower, will cause severe browning and death of embryogenic calli and scales [ 26 ]. However, for the calli of the other two kinds of lilies, coculture for 7 days still maintained a high transformation efficiency, indicating that the tolerance of lily to Agrobacterium may depend on the genotype [ 15 , 27 ]. Drying plant tissue or cells before coculture can also promote T-DNA transfer [ 83 ]. The growth state and speed of calli in dry coculture were better than those in traditional media [ 84 ]. During subsequent resistance screening, only a few tissues were contaminated by the bacterial solution under dry conditions, and the regeneration rate of resistant calli increased significantly [ 34 ]. An appropriate low temperature during coculture also had a positive effect on T-DNA transfer [ 85 , 86 ]. The transformation efficiency of Boehmeria nivea (L.) Gaud. was significantly improved by coculture at 20 °C compared with 15 °C, 25 °C and 28 °C [ 87 ]. In the genetic transformation system of Gossypium hirsutum , 19 °C can significantly increase the regeneration rate of resistant calli and completely inhibit the proliferation of Agrobacterium . However, the effect of temperature on the genetic transformation efficiency of lily has not been clearly reported. In future studies, we can try to optimize the genetic transformation system for lily by adjusting the ambient temperature at each link.

Application of genetic transformation technology

Generation of the crispr/cas9 system.

With the continuous development of gene function research technology, gene modification has been widely used in basic plant research and molecular breeding [ 88 ]. Since CRISPR/Cas9 gene editing technology was successfully applied in Lotus japonicus , because of its simple design and limited operation, this technique has been successfully used in the study of flower anatomy and morphology, flower colour, flowering time, fragrance and stress resistance in various ornamental plants [ 89 , 90 , 91 , 92 ]. Yan et al. [ 26 ] established a stable and efficient genetic transformation system for two lily genotypes using somatic embryos and scales as explants and generated completely albino, light yellow and albino green chimeric mutants via directional knockout of the PDS gene; these authors successfully applied CRISPR/Cas9 technology to lily for the first time. The CRISPR/Cas9 system also validated the feasibility and efficiency of the two genetic transformation systems.

Application for improvement of plant morphogenesis

Morphogenetic genes are key factors that control plant organogenesis and somatic embryogenesis and determine the location of target cells to produce different structures or whole plants [ 8 ]. The functions of many morphogenetic genes have been identified in model plants and important cash crops and have been applied in scientific practice to increase the efficiency of regeneration and genetic transformation [ 93 , 94 ]. In lily, the somatic embryo has always been a good explant for genetic transformation. Plant somatic cells dedifferentiate into embryogenic stem cells under the action of external/internal genetic factors and then divide into somatic embryos. This process is the most critical stage for plant cells to become totipotent [ 95 ]. The most widely used method for somatic embryogenesis (SE) in various plants is the use of exogenous plant growth regulators, especially auxin [ 96 ]. Song et al. [ 17 ] reported that overexpression of LpABCB21 in lily could shorten the time required for SE without changing the exogenous PIC (Picloram). In contrast, the LpABCB21 mutant lines delayed somatic embryo generation by 1–3 days, but the induction rate of adventitious buds was significantly greater than that in the LpABCB21 -overexpressing lines. The study also indicated that the PILS (PIN-LIKES) family member LpPILS7 may participate in auxin regulation through the same mechanism as LpABCB21 , and the somatic embryo induction efficiency of the pils7 mutant was significantly reduced by approximately 10–60%. The importance of miRNAs in SE processes has also been validated in many dicot species and crops [ 82 , 97 ]. In Agrobacterium- mediated Lilium embryo transformation experiments, silencing lpu-miR171a and lpu-miR171b promoted starch accumulation and the expression of key cell cycle genes in calli, significantly accelerated the SE process in Lilium , and resulted in the same phenotype as overexpressing LpSCL6-II and LpSCL6-I . WUSCHEL is a typical gene family involved in the regulation of plant morphogenesis, and its expression is upregulated in many plant SE processes [ 98 , 99 ]. LlWOX9 and LlWOX11 reportedly play a positive regulatory role in the formation of bulbils by influencing cytokinin signalling [ 100 ]. However, the function of WUSCHEL members in Lilium embryogenesis remains to be further verified. It seems that changing the expression level of morphogenetic genes can be an effective means to improve the genetic transformation efficiency of lily, and this topic is worthy of further exploration in the future.

Genetic modification for agronomically important traits

At present, few studies have investigated genetic modification in lily, and most studies have validated the function of target genes only through transient gene transformation (Table  3 ). With the continuous improvement of the genetic transformation system for lily, a few key genes regulating important traits have been identified. In addition to influencing SE processes, many morphogenetic genes are involved in regulating plant organ formation or quality maintenance [ 101 , 102 , 103 ]. LaKNOX1 , a member of the homeobox gene family involved in regulating plant organogenesis, was also further validated in 'Siberia' and 'Sorbonne' [ 18 ]. A recent study revealed that a key gene, LdXERICO , is involved in the regulation of dormancy in Lilium davidii var. unicolour , which indicated that the maintenance of dormancy depends on the ABA-related pathway and that the transcription of LdXERICO is inhibited by the temperature response factor LdICE1 during low-temperature storage, which eventually leads to lily sprouting [ 3 ]. Recently, the LoNFYA7-LoVIL1 module has also been shown to play a key role in orchestrating the phase transition from slow to fast growth in lily bulbs [ 104 ]. Biological and abiotic stresses have a great impact on plant growth and development, and these stresses usually disrupt cellular mechanisms by inducing changes at the physiological, biochemical, and molecular levels in plants [ 105 ]. The identification of key genes involved in the regulation of the stress response in lily was aimed at improving plant resistance to biotic and abiotic stresses. Low temperature, drought, salt stress and abscisic acid treatment can significantly upregulate the expression of LlNAC2 , a member of the NAC transcription factor family. Overexpression of the LlNAC2 gene in tobacco significantly enhances the tolerance of transgenic plants to various abiotic stresses [ 106 ]. Chen et al. [ 18 ] used the genetic transformation system of lily to transform LlNAC2 and successfully generated a transgenic line, which provided favourable support for further clarifying the function of LlNAC2 in coping with abiotic stress in lily species. Typical biological stresses, including bacteria, fungi, viruses, insects and other diseases and pests, seriously negatively affect the quality of ornamental plants [ 89 ]. Several researchers have attempted to increase the resistance of lily plants to pathogens or pests by increasing or decreasing the expression of certain genes. Pratylenchus penetrans (RLN) is one of the main pests and diseases encountered in lily production. The overexpression of the rice cystatin (Oc-IΔD86 ) gene in Lilium longiflorum cv. 'Nellie White' showed that the resistance of transgenic lily to RLN infection was significantly enhanced, and the total nematode population decreased by 75 ± 5%. Compared with wild-type plants, OcIΔD86 -overexpressing plants also exhibited improved growth and development [ 107 ]. Plant resistance to viruses is usually established by transferring the coat protein-encoding gene of the virus into the plant [ 89 ]. Azadi et al. [ 22 ] introduced a cucumber mosaic virus (CMV) replicase defective gene ( CMV2-GDD ) into lily using an Agrobacterium -mediated genetic transformation system and identified two transgenic strains that showed stronger resistance to CMV. In Lilium oriental cv. 'Star Gazer', overexpression of the rice chitinase 10 ( RCH10 ) gene enhanced the resistance of lily to Botrytis elliptica [ 19 ]. Du et al. [ 108 ] identified a gene named LhSorPR4-2 , which encodes a disease course-related protein involved in fighting Botrytis elliptica infection in lily, and the overexpression of LhSorPR4-2 significantly enhanced the resistance of lily to Botrytis . This study also revealed that the function of LhSorPR4-2 was closely related to its chitinase activity. Another study showed that the transcription level of the resistance gene LrPR10-5 was significantly increased in transgenic ‘Siberia’ plants that overexpressed LrWRKY1 , which subsequently promoted resistance to F. oxysporum [ 109 ].

Conclusions

Since the first successful transformation event in lily, remarkable progress has been made; a variety of lily genetic transformation systems have been gradually established, and many excellent new germplasms have been obtained. However, the genetic transformation of lily still faces great challenges due to its strong genotypic dependence. Most related studies have focused on optimizing existing systems, and applicable genetic transformation systems have not yet been established for most lily strains with high market value. For a long time, how to stably and efficiently deliver recombinant gene vectors into plant cells has been the focus of most scholars. At present, the most common delivery method is Agrobacterium -mediated transformation. In addition to the cumbersome tissue culture process, the transformation efficiency also depends greatly on the genotype. The choice of explants for DNA, strains and vectors; culture conditions; and effective selection markers are all major factors that play pivotal roles in successful transformation. At present, most transgenic work in lily is limited by laboratory-scale gene function verification, and even after successful transformation, it is not easy to obtain stable transgenic plants. In recent years, various types of Rhizobium , including Ensifer adhaerens , Ochrobactrum haywardense and Sinorhizobium meliloti , have shown great potential in the transformation of nonagricultural bacterial systems. Some studies have revealed an invisible mechanism for delivering DNA into plant cells, where Sinorhizobium meliloti can infect both monocotyledonous and dicotyledonous plants [ 164 ]. Another way to improve the traditional transformation model is to coexpress developmental regulatory factors or morphogenetic genes during transformation. The overexpression of the developmental regulatory factors GROWTH-REGULATING factor (GRF) and Boby room (Bbm) in maize and sorghum, for which it is difficult to achieve genetic transformation, can significantly improve transformation efficiency [ 101 , 165 ]. Pollen tube transformation is a transformation system that does not require tissue culture, but this method is suitable only for model plants such as Arabidopsis and a few closely related plants. The pollen magnetic transfection-mediated transformation method can be applied to lily, but it may be species- or varietal specific. Zhang et al. [ 166 ] optimized pollen culture conditions, established a new method for the transient transformation of pollen magnetic beads, and concluded that the transformation efficiency was positively correlated with the transverse diameter of pollen and negatively correlated with the ratio of longitudinal diameter to transverse diameter. This study also evaluated the transformation efficiency of Lilium regale L. ‘Sweet Surrender’ and Lilium leucanthum ; L. ‘Sweet Surrender’ and Lilium leucanthum reached 85.80% and 54.47%, respectively, but successful transformation was not achieved in Lilium davidii var. unicolour . Particle bombardment and the electrical shock method are also common methods used in plant genetic transformation. At present, the main explants of the electroshock method are plant protoplasts, but because of their high cost, abundance of chimaeras after transformation and limited stable expression in offspring, these methods cannot be applied to large-scale lily plants. Therefore, exploring genetic transformation systems based on non-tissue culture methods is expected to alleviate the pressure of genetic transformation of lily in the future. Cao et al. [ 167 ] reported a cut-dip-bud (CDB) delivery system. Briefly, the CDB delivery system consists of cutting the junction of plant roots under nonsterile conditions, infecting the upper end with Agrobacterium , taking positive new roots after culture, cutting them into segments and culturing them again to obtain regenerated and transformed plants. Researchers have studied the effects on rubber grass ( Taraxacum koko-saghyz Rodin , TKS), Ipomoea batatas [L.] Lam.), Ailanthus altissima (Mill) Swingle, and Aralia elata (Miq.) The CDB system has been tested in several difficult-to-transform plants, including three woody plants, one of which is Clerodendrum chinense Mabb. The results showed that the CDB delivery system has wide applicability in plant genetic transformation. Furthermore, there is a great need for the validation of promoters other than CaMV35S to achieve optimal expression of transforming genes [ 8 ].

Notably, various plant genetic transformation systems have been further applied to establish RNA interference (RNAi) and gene editing technology systems. To date, there have been few reports on the application of RNAi and CRISPR/Cas9-based gene editing techniques in lilies. In 2019, Yan established a stable and efficient genetic transformation system for somatic embryo regeneration for the first time and successfully conducted targeted gene editing based on CRISPR/Cas9 [ 20 , 26 ]. As one of the sharpest tools in genetic technology, CRISPR/Cas9 “gene scissors” have set off a research boom in basic plant research and directed breeding work. It has great application potential for improving yield, quality, herbicide resistance, abiotic stress resistance and disease resistance. However, there are few successful cases of gene editing using CRISPR/Cas9 technology in lily, which may be related to its high heterozygosity. The stable genetic transformation system has not been widely used, which leads to many difficulties in the study of gene function. With the continuous improvement of the technical system of lily genetic transformation and the emergence of new delivery methods, the combination of multiple transformations may be the only way to develop functional lily genomics in the future, and major breakthroughs in genetic engineering applications in lily breeding are expected not to occur.

Availability of data and materials

The datasets used and/or analysed during the current study available from the corresponding author on reasonable request. No datasets were generated or analysed during the current study.

Abbreviations

Embryogenesis

Acetosyringone

REGULATING factor

Cut-dip-bud

RNA interference

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This work was financed by the National Natural Science Foundation of China (grant number 32302589), the Postdoctoral Science Foundation of China (2023MD744227), Shenyang Innovation Program of Seed Industry (21-110-3-12), Liaoning Province Germplasm Innovation Grain Storage Technology Special Plan (2023JH1/10200010), and the earmarked fund for CARS (CARS-23).

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Xinyue Fan: Writing—original draft. Hongmei Sun: Conceptualization, Writing—review & editing, Supervision, and Funding acquisition. All the authors read and approved the final version of the paper.

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Additional Files 1. Fig. S1: Literature keywords related to the field of lily research that appear together in the map. Keywords that appear more than 30 times are displayed, and different colours represent different cluster.

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Fan, X., Sun, H. Exploring Agrobacterium -mediated genetic transformation methods and its applications in Lilium . Plant Methods 20 , 120 (2024). https://doi.org/10.1186/s13007-024-01246-8

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Published : 09 August 2024

DOI : https://doi.org/10.1186/s13007-024-01246-8

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