Inheritance and expression of pinⅡgene in dh transgenic lines and f_1 hybrids of Chinese cabbage | pinⅡ基因在转基因大白菜DH纯系和试配F1代的遗传及表达

Chino. 摘　　要：马铃薯蛋白酶抑制剂基因-Ⅱ（pinⅡ）是一类控制丝氨酸蛋白酶抑制剂PI-Ⅱ合成的抗虫基因。为了比较pinⅡ基因在转基因大白菜纯系和杂种中的表达特性,为抗虫新品种选育提供理论指导,本实验以转基因大白菜双单倍体纯系（DH系）的5个自交系和与常规育种亲本材料试配的3个F1杂交组合为试材,利用标记性状（除草剂抗性）对试材进行转基因验证,通过小菜蛾饲喂实验对纯系及F1代的离体叶片进行小菜蛾抗性鉴定,同时利用RTq-PCR及蛋白酶抑制活性测定对抗虫性良好的纯系及F1杂种的目的mRNA转录量及目的蛋白合成量进行了测定。结果显示,F1杂种09-2-464对小菜蛾的抑虫效果比纯系T280-2-6强;F1杂种09-2-467对小菜蛾的抑虫效果比纯系T280-3-6强。T180-3-6的自交T2代与杂交F1代对小菜蛾的抑虫作用表现在不同的虫体发育阶段：T280-3-6②主要表现在高致死率和低化蛹率,T280-3-6④和杂种F1代主要表现在降低小菜蛾羽化率上,呈现出抗虫能力上的杂种优势。莲座期T280-3-6②pinⅡmRNA转录量和PI-Ⅱ蛋白表达量分别是F1代09-2-467的5倍和4倍。在植物的不同生长阶段纯系PI-Ⅱ蛋白量均高于F1代。T280-3-6②和F1代09-2-467、09-2-464有希望用于抗虫大白菜育种。

Inglés. Potato protease inhibitors gene-Ⅱ（pinⅡ） is a insect-resistant gene controlling the synthesis of serine protease inhibitors PI-Ⅱ. In order to compare the expression pattern of pinⅡ gene in transgenic cabbage pure lines and hybrids, and provid theoretical guidance for insect resistance breeding sequentially, this experiment used five inbred lines from transgenic cabbage double haploid lines （DH） and three F1 hybrid combinations from DH and conventional breeding materials as test materials, and verified them with herbicide relying on marker trait （herbicides resistance）. we compared DBM （diamondback moth, Plutella xylostella L.） resistance through DBM feeding trials, and quantitated target mRNA and protein by use of RTq-PCR and proteinase inhibitory activity assay for pure lines and F1 hybrids. The results showed that the DBM-inhibitory ability of F1 hybrids 09-2-464 is stronger than T280-2-6, as well, the ability of 09-2-467 is stronger than T280-3-6. The DBM-inhibition of T2 generation and F1 from T180-3-6 act in different growth stage of DBM. T280-3-6② act mainly in the high lethality and lower pupation rate, yet T280-3-6④ and F1 mainly act in reducing DBM feather rate. The quantity of pinⅡ mRNA and PI-Ⅱin T280-3-6② are five and four times of it in 09-2-467 respectively. In all of the plant growth stages, the quantity of PI-Ⅱ in DH is higher than in F1. DH T280-3-6② and F1 09-2-467 and 09-2-464 are hope for insect-resistance transgenic cabbage breeding.