文献类型：期刊文献

中文题名：方竹种胚愈伤组织诱导及遗传转化

英文题名：Embryonic callus induction and genetic transformation of Chimonobambusa quadrangularis

作者：于磊[1] 岳晋军[1] 黄文娟[1] 马漫[1] 张玲[2] 王秋[2] 袁金玲[1]

第一作者：于磊

机构：[1]中国林业科学研究院亚热带林业研究所,杭州311400;[2]四川省宜宾市长宁县林业和竹业局,四川宜宾644300

年份：2022

卷号：58

期号：11

起止页码：2099-2106

中文期刊名：植物生理学报

外文期刊名：Plant Physiology Journal

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家重点研发计划(2021YFD2200505);浙江省农业新品种选育重大专项(2021C02070-4)。

语种：中文

中文关键词：方竹;种胚发育阶段;愈伤组织诱导;组织培养;遗传转化

外文关键词：Chimonobambusa quadrangularis;seed embryo development stage;callus induction;tissue culture;genetic transformation

分类号：S795

摘要：为建立高效的方竹组织培养与遗传转化体系,以方竹种胚为外植体,诱导体细胞胚的发生,研究种胚发育阶段、基本培养基、植物生长调节剂种类及浓度等因素对愈伤组织诱导的影响;选择方竹种胚为受体材料,采用农杆菌介导法,将带有GUS基因的表达载体导入方竹,探讨抗生素浓度、侵染时间和菌液浓度对遗传转化的影响。结果表明:适宜愈伤组织诱导的种胚发育阶段为中Ⅱ期,在添加3.0 mg·L^(-1)2,4-D、1.0 mg·L^(-1)ABA的MS培养基中,愈伤组织诱导率达到87.78%;农杆菌菌液OD600为0.8,超声抽真空侵染20min,GUS基因瞬时表达效率达到86.67%。在添加25 mg·L^(-1)的潮霉素筛选培养基中培养4周后,经PCR检测证明GUS基因已成功转入方竹愈伤组织中,稳定转化效率为25%。本研究结果有助于方竹组培快速繁殖,为利用基因工程技术改良方竹品种提供技术支撑。
In order to establish an efficient tissue culture and genetic transformation system for Chimonobambusa quadrangularis,the seed embryos of Ch.quadrangularis was used as explants to induce somatic embryo formation,the effects of seed embryo development stage,basic medium,plant growth regulator type and concentration callus induction were studied.The Ch.quadrangularis seed embryos were selected as materials,and a binary vector withβ-glucuronidase(GUS)was transferred to the callus of Ch.quadrangularis by Agrobacterium-mediated transformation,the effects of antibiotic concentration,infection time and Agrobacterium concentration on genetic transformation were studied.The results showed that the suitable seed embryo development stage for callus induction was the middle stageⅡ.The callus induction rate was 87.78%at MS+3.0 mg·L^(-1)2,4-D+1.0 mg·L^(-1)ABA.When the OD600 of Agrobacterium concentration was 0.8,after ultrasonic vacuum infection for 20 min,the transient expression efficiency of GUS gene reached 86.67%.After culturing in the selection medium supplemented with 25 mg·L^(-1)of hygromycin for 4weeks,PCR detection showed that the GUS gene has been successfully transferred into Ch.quadrangularis callus,and the stable transformation efficiency is 25%.The results of this study are helpful to the rapid propagation of Ch.quadrangularis by tissue culture and provide technical support for variety improvement by genetic engineering technology.