偏穗鹅观草胚轴愈伤组织诱导体系建立  

Establishment of Hypocotyl Callus Induction System of Roegneria komarovi(i Nevski)Nevski

作　　者：金丹青 顾鹏程 王玉杰 胡尚钦 汪军成[2,3] 马小乐[2,3] 司二静[2,3] 姚立蓉 李葆春[1,2] JIN Danqing;GU Pengcheng;WANG Yujie;HU Shangqin;WANG Juncheng;MA Xiaole;SI Erjing;YAO Lirong;LI Baochun(College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;State Key Laboratory of Aridland Crop Science/Gansu Provincial Key Laboratory of Crop Improvement&Germplasm Enhancement,Lanzhou 730070,China;College of Agronomy,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学生命科学技术学院,甘肃兰州730070 [2]省部共建干旱生境作物学国家重点实验室/甘肃省作物遗传改良与种质创新重点实验室,甘肃兰州730070 [3]甘肃农业大学农学院,甘肃兰州730070

出　　处：《山西农业科学》2022年第9期1215-1220,共6页Journal of Shanxi Agricultural Sciences

基　　金：国家自然科学基金项目(31860377);甘肃省自然科学基金重点项目(21JR7RA801);国家现代农业产业技术体系(CARS-05-02A-02)。

摘　　要：偏穗鹅观草具有抗病、抗旱、耐盐等有利性状,是小麦主要的二级基因源,将偏穗鹅观草的有利基因引入小麦,创造优异变异材料至关重要。通过建立偏穗鹅观草愈伤组织诱导体系,成功诱导出高质量愈伤组织进行继代培养,可为小麦和偏穗鹅观草原生质体融合提供技术支持,从而为小麦新种质创制和抗性育种奠定基础。对偏穗鹅观草不同部位外植体进行愈伤组织诱导,通过使用不同激素组合的培养基,筛选出适合偏穗鹅观草愈伤组织诱导的外植体及激素组合,以诱导效果较好的外植体为材料,进一步通过正交试验筛选适合其愈伤诱导的最佳培养基,为偏穗鹅观草基因的利用创造条件。结果表明,用胚轴进行愈伤组织的诱导优于叶柄和叶片;2,4-D能够显著影响偏穗鹅观草愈伤组织的诱导率,对诱导偏穗鹅观草胚轴形成愈伤影响最大,6-BA和NAA的作用次之。由正交试验诱导率的极差分析结果可知,最优组合为A1B2C3,即0.5 mg/L 6-BA、1.0 mg/L 2,4-D、0.6 mg/L NAA。胚轴为最佳外植体,2,4-D对外植体诱导愈伤起着关键作用,胚轴诱导愈伤的最佳培养基为MS+5.0 mg/L 2,4-D+0.1 g肌醇+30 g/L蔗糖+5 g琼脂(pH=5.8),诱导率可达到85%~90%。Roegneria komarovii(Nevski)Nevski is the main secondary gene source of wheat with favorable traits such as disease resistance,drought resistance,and salt tolerance.It is very important to introduce elite genes of Roegneria komarovii(Nevski)Nevski into wheat to create excellent variant materials.By establishing a callus induction system of Roegneria komarovii(Nevski)Nevski,high quality callus is succsessfully induced for subculture,which can provide technical support for protoplast fusion of wheat and Roegneria komarovii(Nevski)Nevski,laying foundation for germplasm creation and resistance breeding of wheat.In this study,by callus induction in explants of different parts of Roegneria komarovii(Nevski)Nevski and different concentrations of hormone,the optimal explant and hormone combinations suitable for callus induction of Roegneria komarovii(Nevski)Nevski were screened.The explants with excellent induction effect were used for further orthogonal test to screen the optimum medium suitable for the callus induction,so as to create conditions for the utilization of Roegneria komarovii(Nevski)Nevski gene.The results showed that the callus induction with hypocotyl was better than that of petiole and leaf.2,4-D could significantly affect induction rate of callus of Roegneria komarovii(Nevski)Nevski,and had the greatest effect on inducing callus from the hypocotyl of Roegneria komarovii(Nevski)Nevski,followed by 6-BA and NAA.The orthogonal test indicated that the optimal hormone combination for callus induction was A1B2C3,namely 0.5 mg/L of 6-BA,1.0 mg/L of 2,4-D,0.6 mg/L of NAA.Hypocotyl was the best explant,and 2,4-D played a key role in callus induction.The optimal medium for callus induction with hypocotyl was MS+5.0 mg/L of 2,4-D+0.1 g of inositol+30 g/L of sucrose+5 g of agar(pH=5.8),its induction rate reached to 85%-90%.

关 键 词：偏穗鹅观草 愈伤组织诱导 正交设计 胚轴 

分 类 号：S512.1[农业科学—作物学]