'双丰1号'菜豆离体再生体系的建立OA
Establishment of in Vitro Regeneration System of'Shuang Feng No.1'Common Bean(Phaseolus vulgaris)
菜豆(Phaseolus vulgaris)为重要豆科蔬菜,离体再生困难限制了其遗传转化与分子育种效率.为建立高效、稳定的菜豆离体再生体系,以'双丰一号'菜豆为材料,以子叶节、下胚轴、真叶、子叶为外植体,采用直接再生(愈伤途径)与间接再生(子叶节直接出芽)2种方式优化植物生长调节剂配比、低温及高渗逆境条件,系统筛选愈伤诱导、胚性愈伤分化、不定芽发生及生根移栽的最佳参数,建立菜豆离体再生体系.结果显示:(1)松散型愈伤诱导最佳外植体为下胚轴、子叶节,最佳培养基为MS+1 mg/L 2,4-D,诱导率分别达93.33%、90.00%.(2)菜豆胚性愈伤最佳诱导培养基为MS+0.5 mg/L 2,4-D+0.25 mg/L 6-BA,4℃低温预处理2～5 d可显著促进胚性化,但未能实现稳定增殖与植株再生.(3)间接再生以子叶节为最佳外植体,最佳诱导培养基为MS+2 mg/L 6-BA,不定芽诱导率93.33%,单外植体平均出芽6.45个;TDZ虽诱导率100%,但易产生畸形芽与玻璃化苗.(4)最优生根培养基为1/2 MS+1 mg/L IBA,生根率96.67%,炼苗移栽成活率84.16%,再生植株可正常开花结荚.本研究成功建立了'双丰1号'菜豆高效离体再生体系,可为菜豆离体快繁、种质创新及遗传转化体系构建提供技术支撑,后续可进一步优化胚性愈伤增殖与体胚发生条件,完善全流程再生系统.
Common bean(Phaseolus vulgaris)is an important legume vegetable,and its in vitro regeneration is difficult,which limits the efficiency of genetic transformation and molecular breeding.In order to establish an efficient and stable in vitro regeneration system of common bean,this study used'Shuangfeng No.1'common bean as material,cotyledonary nodes,hypocotyls,true leaves and cotyledons as explants,and used direct regeneration(callus pathway)and indirect regeneration(cotyledonary nodes directly bud)to optimize the ratio of plant growth regulators,low temperature and high osmotic stress conditions.The optimal parameters of callus induction,embryogenic callus differentiation,adventitious bud formation and rooting transplanting were systematically screened,and the in vitro regeneration system of common bean was established.The results showed that:(1)the best explants for loose callus induction were hypocotyls and cotyledonary nodes.The best medium was MS+1 mg/L 2,4-D,and the induction rates were 93.33%and 90.00%,respectively.(2)The optimal medium for inducing embryogenic callus in common bean(Phaseolus vulgaris)was MS+0.5 mg/L 2,4-D+0.25 mg/L 6-BA.Embryogenic callus was more easily induced when the explants were cultured at low temperature for 2 to 5 days.However,successful regeneration through callus was not achieved in this experiment,and further research is needed.(3)The indirect regeneration experiment indicated that the best explant was the cotyledon node,and the optimal induction medium was MS+2 mg/L 6-BA,with an adventitious bud induction rate of 100%.(4)The induced adventitious buds could be transferred to 1/2 MS+1 mg/L IBA rooting medium to regenerate complete plants.The rooting rate was 96.67%,the survival rate of transplanted seedlings was 84.16%,and the regenerated plants could flower and pod normally.This experiment established an efficient in vitro regeneration culture system for common bean(Phaseolus vulgaris),enabling rapid propagation of plants in vitro and laying the foundation for the establishment of a genetic transformation system for common bean(Phaseolus vulgaris).
许莹楠;刘艳军;黄俊轩;韩启厚;高逸卓;古瑜
天津农学院园艺园林学院,天津 300392天津农学院园艺园林学院,天津 300392天津农学院园艺园林学院,天津 300392天津市农业科学院/蔬菜生物育种全国重点实验室,天津 300192||天津科润农业科技股份有限公司蔬菜研究所/天津市蔬菜遗传育种企业重点实验室,天津 300384天津农学院园艺园林学院,天津 300392天津市农业科学院/蔬菜生物育种全国重点实验室,天津 300192||天津科润农业科技股份有限公司蔬菜研究所/天津市蔬菜遗传育种企业重点实验室,天津 300384
农业科技
菜豆松散型愈伤组织胚性愈伤组织子叶节再生不定芽植物生长调节剂
common bean(Phaseolus vulgaris)loose callusembryogenic calluscotyledons regenerationadventitious budsplant growth regulators
《中国农学通报》 2026 (10)
52-60,9
天津市农业科学院财政种业创新研究项目天津市科技重大专项与工程计划项目"荚用菜豆新种质创制方法及种质资源创新研究"(17ZXZYNC00030).
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