油莎豆原生质体制备的组织筛选及条件优化  

Tissue Screening and Condition Optimization of Protoplast Preparation of Cyperus esculentus

作　　者：张向歌 朱雅婧 陆莉莉[1] 宋万献 张新友[1] 胡海燕[1] 于美琴 李居政 王会伟[1] ZHANG Xiangge;ZHU Yajing;LU Lii;SONG Wanxian;ZHANG Xinyou;HU Haiyan;YU Meiqin;LI Juzheng;WANG Huiwei(Industrial Crop Research Institute,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)

机构地区：[1]河南省农业科学院经济作物研究所,河南郑州450002

出　　处：《河南农业科学》2023年第12期49-56,共8页Journal of Henan Agricultural Sciences

基　　金：河南省重大科技专项(211100110100);河南省农业科学院优秀青年科技基金项目(2022YQ19);河南省重点研发与推广专项(科技攻关)项目(232102110261);河南省农业科学院自主创新项目(2023ZC018)。

摘　　要：为高效获得油莎豆原生质体,采用单因素试验方法进行油莎豆原生质体制备的组织部位(幼芽、幼根、幼叶、幼叶鞘、分蘖节和匍匐茎)筛选以及酶解条件(酶组合用量、酶解时间、渗透压稳定剂用量和酶液pH值)优化。结果表明,油莎豆原生质体制备的最佳组织部位为幼嫩的分蘖节;最优酶解条件:酶组合用量为2.0%纤维素酶+0.9%离析酶、酶解时间为6h、渗透压稳定剂甘露醇用量为11%、酶液pH值为6.0。此条件下,0.2 g油莎豆幼嫩分蘖节加至4 mL酶液中,原生质体的产量可达22.91×10^(5)个/g,活力可达90.72%。综上,获得了高效制备油莎豆原生质体的方法,可为后续油莎豆原生质体融合、种质创新以及杂交育种奠定基础。In order to obtain Cyperus esculentus protoplasts efficiently,the tissue parts including young shoots,young roots,young leaves,young leaf sheaths,tillering nodes and stolons were screened for protoplast preparation by single factor test method,and the enzymatic hydrolysis conditions such as enzyme combination dosage,enzymatic hydrolysis time,osmotic pressure stabilizer dosage and pH value of the enzyme solution were also optimized.The results showed that the best tissue part for protoplast preparation of Cyperus esculentus was young tillering node;and the optimal enzymatic hydrolysis conditions were as follows:the enzyme combination dosage was 2.0%cellulase+0.9%macerozyme,the enzymatic hydrolysis time was 6 h,the osmotic pressure stabilizer(mannitol)dosage was 11%,and the pH value of the enzyme solution was 6.0.Under these conditions,0.2 g of Cyperus esculentus tillering nodes were added to 4 mL of enzyme solution,and then the protoplast yield could reach 22.91×10^(5)per gram and the protoplast activity could reach 90.72%.In this study,an efficient method for protoplast preparation of Cyperus esculentus was obtained,which could lay a foundation for subsequent protoplast fusion,germplasm innovation and hybrid breeding of Cyperus esculentus.

关 键 词：油莎豆 原生质体制备 组织筛选 酶解条件 优化 种质创新 

分 类 号：S565.9[农业科学—作物学]