紫花苜蓿胚性愈伤组织诱导及胚状体分化研究  

Study on Embryogenic Callus Induction and Embryoid Differentiation of Alfalfa

作　　者：王晓春 高婷 杨炜迪 王川 陈彩锦 Wang Xiaochun;Gao Ting;Yang Weidi;Wang Chuan;Chen Caijin(Institute of Animal Sciences,Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan 750000,Ningxia,China;Guyuan Branch of Ningxia Academy of Agriculture and Forestry Sciences,Guyuan 756000,Ningxia,China)

机构地区：[1]宁夏农林科学院动物科学研究所,宁夏银川750000 [2]宁夏农林科学院固原分院,宁夏固原756000

出　　处：《作物杂志》2023年第4期98-103,共6页Crops

基　　金：宁夏自然科学基金(2020AAC03326,2021AAC03285);宁夏回族自治区政府农作物育种重大专项(2014NYZ03)。

摘　　要：以苜蓿(固原紫花)下胚轴、茎段和叶片为外植体,开展了胚性愈伤组织的诱导及分化研究。结果表明,2,4-二氯苯氧乙酸(2,4-D)浓度为2和4mg/L时,愈伤组织诱导率为100.0%,浓度升为6和8mg/L时,诱导率逐渐降低,说明高浓度的2,4-D对苜蓿外植体愈伤组织诱导有抑制作用。MS培养基添加2,4-D(1.0、2.0、3.0、4.0mg/L),并配合0.5mg/L 6-BA(6-苄基氨基嘌呤)或KT(激动素)后,3种外植体的诱导率均为100%,24d内能完全脱分化,且在质地和形态上无差别。转入分化培养基1/2 MS后,所有配方都能分化出胚状体,2,4-D浓度2.0mg/L、KT浓度0.5mg/L组合的分化率最高(12.6%)。改进的分化培养基(1/2 MS+0.4mg/L KT)分化率提高到42.7%。以诱导出的再生苗叶片和叶柄为外植体,发现体细胞胚胎诱导能力高度遗传,在添加2,4-D(1.0~8.0mg/L)组合0.5mg/L 6-BA或KT的培养基内都能100%诱导出大量胚状体并发育为完整植株。由此可见,通过再生植株可建立紫花苜蓿高频再生体系。The induction and differentiation of embryogenic callus were studied using hypocotyls,stem segments and leaves of Guyuan alfalfa as explants.The results showed that the callus induction rate was 100%when the concentrations of 2,4-D was 2 and 4mg/L.When the concentration of 2,4-D added to 6 or 8mg/L,the induction rate gradually decreased,indicating that high concentration of 2,4-D could inhibit the callus induction.The induction rates were 100%when the concentrations of 2,4-D(1.0,2.0,3.0,4.0mg/L)combined with 0.5mg/L 6-BA or KT using three kinds of explants cultured on MS medium.The callus could dedifferentiate completely within 24 days,and there was no difference in texture and morphology.Embryoids could be differentiated from all the formulations in the differentiation medium of 1/2 MS,but the highest differentiation rate was only 12.6%when the concentration of 2,4-D was 2.0mg/L combined with 0.5mg/L KT.The differentiation rate was increased to 42.7%by supplementing 0.4mg/L KT in differentiation medium.Using the leaves and petioles of the regenerated plantlets as explants,the embryoids could be easy to induced and developed into strong plants in the medium supplemented with 2,4-D(1.0-8.0mg/L)and 0.5mg/L 6-BA or KT because the inducible ability of somatic embryoid was highly inherited.In conclusion,the high frequency regeneration system of alfalfa could be established by regenerating plants.

关 键 词：紫花苜蓿 愈伤组织 体细胞胚 再生植株 

分 类 号：S541.9[农业科学—作物学]