菠菜(Spinacia oleracea L)是研究雌雄异株植物性别分化的模式材料,其性别分化不但受到性别决定基因的控制,还受到环境条件的影响,干旱是其中的一个重要因素,但是关于聚乙二醇(Polyethylene glycol,PEG)模拟土壤干旱环境对菠菜性别分化影响的分子机制目前尚未见报道。试验以日本大叶菠菜(S.oleracea cv.Japanese leaves)为供试材料,利用不同浓度的PEG6000进行处理,研究干旱环境对菠菜性别分化的影响。采用温室盆栽种植方式,分别用5%、10%的PEG6000模拟土壤干旱环境对盆栽菠菜处理72 h,处理株数均为100株,并同时设置对照组。结果表明,与对照组雌雄比例(0.96)相比,PEG60005%处理组的雌雄比例(1.09)与10%处理组(0.97)均接近于对照组;对发生性反转数进行统计,结果表明,PEG6000 10%处理组低于5%处理组;利用菠菜Y染色体紧密连锁的雄性特异标记T11A对处理前、后的植株进行鉴定,发现菠菜植株出现了性反转现象,且均为雌株转变为雄株。琼脂糖凝胶电泳结果显示,性反转后,雄性特异条带也随之出现。试验首次对PEG6000模拟土壤干旱环境对菠菜性别分化影响的分子机制进行了探究,其研究结果对于进一步揭示菠菜性别分化机理具有重要意义。
Spinacia oleracea L is the model material for studying the sex differentiation of dioecious plants. Sex differentiation of dioecious plants was controlled not only by the sex-determining gene but also affected by environmental conditions, of which drought is one of the important factors. However, the molecular mechanism of the effect of drought stress on S. oleracea sex differentiation under polyethylene glyco(PEG) treatment has not been reported. Taking S. oleracea cv. Japanese leaves as material, treating with different concentration of PEG6000, the effect of drought stress on S. oleracea gender differentiation was studied. Potting plants in greenhouse, 200 plants were treated by 5% and 10% PEG6000 respectively while the control group was treated using water. After 72 h, the male and female ratio of treated groups was compared with that of control group(0.96). the results showed that the female-to-male ratio of 5% PEG6000 treatment(1.09) and 10% PEG6000treatment(0.97) are close to that of the control group. The statistics showed that the sex-reversal number of 10% PEG6000 treatment was lower than that of 5% PEG6000 treatment. Using the Y chromosome linked male specific molecular markers T11 A to identify the S. oleracea sex phenomenon before and after PEG6000 treatment, the results showed that the female-tomale sex reversal phenomenon happened. Agarose gel electrophoresis results showed that after female-to-male sex reversal,male specific bands also emerged. The molecular mechanism of the effect of drought stress on spinach sex differentiation under PEG6000 treatment were studied for the first time, which has great significance for further inquiry to the S. oleracea gender differentiation mechanism.