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In vitro regeneration of Phaseolus vulgaris L. via direct and indirect organogenesis
Yu Yan,Liu Dajun,Liu Chang,Yan Zhishan,Yang Xiaoxu,Feng Guojun 한국식물생명공학회 2021 Plant biotechnology reports Vol.15 No.3
A stable and efcient regeneration system is a necessary condition for the genetic transformation of Phaseolus vulgaris L. In this study, cotyledonary nodes with one cotyledon, cotyledonary nodes with two cotyledons, embryonic axes and root segments of 5-day-old germinated seeds were used as explants to evaluate explants to induce shoots and calli. Besides, using cotyledonary nodes with one cotyledon as explants, the efects of diferent concentrations of auxins and cytokinins on the direct and indirect organogenesis of common bean were investigated. As a result, embryonic axes and cotyledonary nodes with two cotyledons were the most efective explant types for the formation of shoots and calli. The cotyledonary nodes with one cotyledon were cultured in the shoot induction medium containing 7 mg L−1 6-benzyladenine (BAP) and 0.2 mg L−1 α-naphthaleneacetic acid (NAA). 74.21% of explants directly formed shoots and average number of shoots was up to 2.91. On the rooting medium supplemented with 0.5 mg L−1 indole-3-butyric acid (IBA), the rooting rate of shoots reached 100% and the number of main roots formed was the most. The addition of 0.5 mg L−1 thidiazuron (TDZ) was optimum for callus induction, and the induction rate could reach 91.51%. The shoot regeneration frequency was 93.33% when calli subcultured on the shoot induction medium supplemented with 1.0 mg L−1 BAP. Average 4.27 shoots were produced by per callus and rooted on the shoot induction medium. This report descripts an efcient common bean regeneration system through direct and indirect organogenesis, which can provide reference for its subsequent genetic transformation research.
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Chang Liu,Xiaoxu Yang,Zhishan Yan,Youjun Fan,Guojun Feng,Dajun Liu 한국유전학회 2019 Genes & Genomics Vol.41 No.12
and quality. At present, little is known about the genes and molecular regulation mechanism in cold response in snap bean exposed to low temperature. Objectives Our objectives were to identify the low temperature response genes in snap bean and to examine differences in the gene response between cold-tolerant and cold-sensitive genotypes. Methods We used two highly inbred snap bean lines in this study, the cold-tolerant line ‘120’, and the cold-sensitive line ‘093’. The plants were grown to the three leaf and one heart stage and exposed to 4 °C low temperature. We used RNA sequencing (RNA-seq) to analyze the differences of gene expression. Results 988 and 874 cold-responsive genes were identified in ‘T120 vs CK120’ and ‘T093 vs CK093’ (‘T’ stands for low temperature treatment, and ‘CK’ stands for control at room temperature), respectively. Of these, 555 and 442 genes were unique to cold-stressed lines ‘120’ and ‘093’, respectively compared to the control. Our analysis of these differentially expressed genes indicates that Ca2+, ROS, and hormones act as signaling molecules that play important roles in low temperature response in P. vulgaris. Altering the expression of genes in these signaling pathways activates expression of downstream response genes which can interact with other signaling regulatory networks. This may maintained the balance of ROS and hormones, making line ‘120’ more cold-tolerant than line ‘093’. Conclusion Our results provide a preliminarily understanding of the molecular basis of low temperature response in snap bean, and also establish a foundation for the future genetic improvement of cold sensitivity in snap bean by incorporating genes for cold tolerance.
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Optimization of an indirect regeneration system for common bean (Phaseolus vulgaris L.)
Xiong Luxi,Liu Chang,Liu Dajun,Yan Zhishan,Yang Xiaoxu,Feng Guojun 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.6
The main problems associated with growing common bean (Phaseolus vulgaris L.) callus are low proliferation and differentiation, and high browning. In this study, common bean callus induced by cotyledon nodes was used as explant material to investigate the effects of different exogenous substances on callus regeneration, as well as the correlation between callus browning and the changes in superoxide dismutase (SOD), peroxidase (POD), and polyphenol oxidase (PPO) enzyme activities during callus culture. Adding AgNO3, CoCl2, Putrescine (Put), and 1-aminocyclopropane-1-carboxylic acid (ACC) to the callus growth medium at appropriate concentrations could significantly improve callus proliferation and differentiation efficiency, while also reducing the degree of browning, compared to controls. Callus explants were cultured in callus proliferation and differentiation medium (CPADM) containing 5 mg·L−1 AgNO3, 15 mg·L−1 Put, 5 mg·L−1 CoCl2, or 0.02 mM ACC, with optimal callus growth at these dosages. After treatment with the four exogenous substances, callus browning was inversely correlated with SOD activity but positively correlated with POD and PPO activities. The maximum rooting frequency of shoots was observed when rooting media was treated with 1 mg·L−1 Indole butyric acid (IBA) or 0.1 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D). The average number of primary roots, root length, and root fresh weight were higher after these treatments. The indirect regeneration issue was largely resolved for common bean by the callus culture technique applied in this study, which provides a framework for genetic modification, germplasm preservation, and bean application.
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