白花玉石籽石榴遗传转化体系的建立  

Establishment of genetic transformation system for Baihuayushizi pome-granate

作　　者：唐靖雯 王宁 伍程程 王麒麟 丁文豪 葛伟强 郭宁 钱晶晶 TANG Jingwen;WANG Ning;WU Chengcheng;WANG Qilin;DING Wenhao;GE Weiqiang;GUO Ning;QIAN Jingjing(College of Agriculture,Anhui Science and Technology University,Fengyang 233100,Anhui,China;Anhui Zhongyi Agricultural Sci-ence&Technology Co.,Ltd.,Huaiyuan 233400,Anhui,China;Agricultural Technology Extension Center of Jingshan City,Jingmen City,Hubei Province,Jingshan 431800,Hubei,China)

机构地区：[1]安徽科技学院农学院,安徽凤阳233100 [2]安徽中以农业科技有限公司,安徽怀远233400 [3]湖北省荆门市京山市农业技术推广中心,湖北京山431800

出　　处：《果树学报》2024年第12期2621-2633,共13页Journal of Fruit Science

基　　金：安徽省高校自然科学研究重大项目(2023AH040281);安徽中以农业科技有限公司产学研合作项目(AKZY2022110)。

摘　　要：【目的】为更好地研究石榴的基因功能,以白花玉石籽石榴为试材,构建出一种农杆菌介导的石榴遗传转化体系。【方法】在已有的石榴组织培养体系的基础上,通过筛选不同外植体和优化激素组合,获得石榴最佳再分化途径;探究农杆菌介导法中抗生素浓度、预培养时间、菌液浓度、侵染时间和抑菌剂浸洗时间对白花玉石籽石榴遗传转化效率的影响。【结果】终质量浓度为0.22 mg·L^(-1)6-BA和0.60 mg·L^(-1) IBA的WPM培养基能显著提高石榴外植体的再分化率,其中嫩茎分化率达96.30%±5.20%;50 mg·L^(-1)卡那霉素和200 mg·L^(-1)特美汀是筛选抗性芽的最佳质量浓度;白花玉石籽石榴遗传转化的最适组合为:预培养3 d、菌液OD600=0.8、侵染10 min、200 mg·L^(-1)特美汀浸洗15 min。最后,通过检测GFP荧光,验证上述遗传转化体系所获得的植株,阳性率为26.00%。【结论】成功建立了农杆菌介导的白花玉石籽石榴嫩茎遗传转化体系,为石榴基因功能验证提供有力的技术支持。【Objective】Pomegranate is a favorate among consumers because of its high economic,nutri-tional and medicinal values.The rapid development of molecular biology has made genetic transforma-tion an important means to obtain new excellent germplasm of crops and carry out gene function verifi-cation,as well as apply an effective supplement to traditional breeding.At present,the genetic transfor-mation system of pomegranate is incomplete,resulting in the lagging of gene function research and mo-lecular breeding in pomegranate compared with other fruit crops.This experiment aimed to build a sta-ble and efficient genetic transformation system for Baihuayushizi pomegranate,with a view to provid-ing satisfactory technical support for gene function verification of pomegranate and the improvement of its germplasm resources.【Methods】In this study,sterile seedlings of Baihuayushizi pomegranate were used.On the basis of the tissue culture system in early stage,the redifferentiation system of pomegran-ate and the concentrations of kanamycin and timentin were screened,followed by a discussion on the re-lated influencing factors concerning genetic transformation such as pre-culture time,concentration of Agrobacterium,infection time and socking time of antibacterial agents.Finally,the optimal genetic transformation system for Baihuayushizi pomegranate mediated by Agrobacterium was established.【Results】The addition of 0.22 mg·L^(-1)6-BA and 0.60 mg·L^(-1) IBA into the WPM medium significantly improved the redifferentiation of pomegranate implants,which was manifested as higher differentiation rates of leaves and tender stems compared to other treatment portfolios.The differentiation rates were 83.93%±2.52%and 96.30%±5.20%,respectively,and the differentiation rate of tender stems increased by 14.74%compared with that of leaves.In addition,tender stems with a high differentiation rate were used as the receptor,and both induction rate and differentiation rate were significantly higher than other treatments without addit

关 键 词：石榴 再生途径优化 农杆菌介导法 遗传转化体系 

分 类 号：S665.4[农业科学—果树学]