农杆菌介导转化玉米自交系B104体系的优化  

Optimization of Agrobacterium-mediated Transformation System of Maize Inbred Line B104

作　　者：潘梦影 王寒 李亚永 朱俊杰 徐秋月 纪坤 石慧玥 张鑫悦 董永彬[1] 李玉玲[1] Pan Mengying;Wang Han;Li Yayong;Zhu Junjie;Xu Qiuyue;Ji Kun;Shi Huiyue;Zhang Xinyue;Dong Yongbin;Li Yuling(College of Agronomy,Henan Agricultural University,Zhengzhou,450046)

机构地区：[1]河南农业大学农学院,郑州450046

出　　处：《分子植物育种》2024年第20期6731-6739,共9页Molecular Plant Breeding

基　　金：国家留学基金项目(201708410061);国家自然科学基金项目(31671770);河南省自然科学基金项目(212300-410356);河南省玉米产业技术体系(HARS-22-02-S)共同资助。

摘　　要：为了提高玉米遗传转化效率,本研究以玉米自交系B104幼胚为受体,β-葡萄糖苷酸酶基因(β-glucuronidase,GUS)为报告基因,以GUS基因的瞬时表达率为指标,通过不同温度、不同相对离心力(relative centrifuge force,RCF)和共培养培养基不同MS盐浓度处理玉米幼胚,观察统计GUS基因的瞬时表达效率以及愈伤组织诱导形成的影响;培养基中添加不同浓度的硝酸银、6-BA处理玉米幼胚和愈伤组织,观察对愈伤组织形成和再生成苗影响。结果表明:(1)玉米幼胚热激预处理温度为43℃时,GUS基因瞬时表达效率最好,且愈伤组织形成率为100%;(2)10000 RCF处理玉米幼胚,对盾片区GUS瞬时表达率和愈伤组织诱导形成率具有作用,能有效提高玉米幼胚的遗传转化效率;(3)共培养培养基50%MS盐浓度不影响愈伤组织的形成,同时GUS瞬时表达情况较好;(4)激素6-BAP浓度对玉米愈伤组织发育和再生有明显的促进作用;(5)硝酸银对玉米幼胚愈伤组织的形成有明显的抑制作用,但对再生成苗有显著的促进作用。本研究为进一步优化玉米自交系的遗传转化效率提供了科学依据。To improve the efficiency of maize genetic transformation,here,the young embryos of maize inbred line B104 were used as the receptor,β-glucuronidase gene(GUS)was used as the reporter gene,and the transient expression rate of GUS gene as the index.The transient expression efficiency of GUS gene and its effect of callus induction were observed by treating maize young embryos with different temperatures,different relative centrifuge force(RCF)and different MS salt concentrations in co-culture medium;the callus formation and seedling formation were observed in maize young embryos and callus tissue treated with different concentrations of silver nitrate and 6-BA,respectively.The results showed that:(1)When the heat shock pretreatment temperature was 43℃,the transient expression efficiency of GUS gene could reach the best,and the callus formation rate was 100%;(2)10000 RCF treatment had effects on GUS transient expression rate and callus induction rate in the scutellum region,which could effectively improve the genetic transformation efficiency of maize young embryos;(3)The formation of callus was not affected by 50%MS salt concentration in co-culture medium,and the transient expression of GUS was better;(4)The hormone content of 6-BAP significantly promoted callus formation and seedling regeneration;(5)Silver nitrate significantly inhibited callus formation in young maize embryos,but significantly promoted seedling formation.This study provided a scientific basis for further optimizing the genetic transformation efficiency of maize inbred lines.

关 键 词：玉米 自交系 农杆菌介导法 GUS染色 

分 类 号：S513[农业科学—作物学]