紫花苜蓿胚性愈伤组织诱导与增殖的研究

Induction and Proliferation of Embryogenic Callus of Alfalfa

为在不同栽培苜蓿(Medicago sativa)品种上,建立稳定、快速且高效的组织培养体系,本试验以品种‘游客’、‘Paola’和‘Sardi7’为研究材料,探究了苜蓿外植体、激素配比及金属离子(Cu 2+、Ag+),对其胚性愈伤组织的诱导效果。结果表明:下胚轴为诱导愈伤组织的最佳外植体;MS+1 mg·L^(-1)2,4-D+0.2 mg·L^(-1)KT为培养基时‘游客’和‘Paola’胚性愈伤诱导率分别为24.7%,16.7%;MS+2 mg·L^(-1)2,4-D+0.2 mg·L^(-1)KT为培养基时‘Sardi7’胚性愈伤诱导效果最佳为27.3%;1μmol·L^(-1)和5μmol·L^(-1)Cu^(2+)均能显著促进‘Sardi7’‘游客’和‘Paola’愈伤组织的诱导;5μmol·L^(-1)Ag^(+)显著诱导‘游客’和‘Paola’形成胚性愈伤组织,诱导率分别提高25.3%,46.7%(P<0.05)。本研究探索了诱导苜蓿愈伤组织的最佳外植体及其培养条件、金属离子促进不同苜蓿品种再生的作用,为突破同源四倍体栽培苜蓿品种基因型上的限制、提高胚性愈伤组织出愈率提供了思路。

In order to establish a stable,rapid and efficient tissue culture system on different cultivated alfalfa(Medicago sativa)varieties,this experiment took the varieties‘Tourist’‘Paola’and‘Sardi7’as the research materials,investigated the effects of alfalfa explants,hormones ratio and metal ions(Cu^(2+),Ag^(+))on induction of embryonic callus.The results showed that hypocotyl was the best explants to induce callus.When MS+1 mg·L^(-1)2,4-D+0.2 mg·L^(-1)KT were cultured,embryogenic callus induction rates of‘Tourist’and‘Paola’were 24.7%and 16.7%respectively.And the embryogenic callus induction effect of‘Sardi7’was 27.3%when MS+2 mg·L^(-1)2,4-D+0.2 mg·L^(-1)KT were cultured.1μmol·L^(-1)and 5μmol·L^(-1)Cu^(2+)could promote callus induction of‘Sardi7’,‘Tourist’and‘Paola’significantly.5μmol·L^(-1)Ag^(+)significantly induced embryogenic callus of‘Tourist’and‘Paola’,and the induction rates increased by 25.3%and 46.7%,respectively(P<0.05).In this study,we explored the best explants to induce alfalfa callus and their culture conditions,as well as the role of metal ions in promoting regeneration of different alfalfa varieties,which provided a solution for breaking through the genotypic restriction of autotetraploid cultivated alfalfa varieties and improving the healing rate of embryonic callus.