小麦生理型雄性不育系中天冬氨酸蛋白酶与绒毡层代谢的相关性分析  

The Research on the Correlation Between Aspartic Proteinase and Anther Tapetum in Physiological Male Sterility Wheat

作　　者：巨岚 朱启迪[1] 张改生[1] 张姣[1] 于永昂[1] 刘红占[1,2] 牛娜 王军卫[1] 

机构地区：[1]西北农林科技大学/国家杨凌农业生物技术育种中心/国家小麦改良中心杨凌分中心/小麦育种教育部工程研究中心/陕西省作物杂种优势研究与利用重点实验室,陕西杨凌712100 [2]周口师范学院生命科学与农学学院,河南周口466001

出　　处：《核农学报》2018年第3期430-437,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家高技术研究发展计划重大专项(863计划)(2011AA10A106);国家科技支撑计划(2015BAD27B01);国家自然科学基金(31171611;31371697);陕西省科技统筹创新工程计划课题(2014KTZB02-01-02)

摘　　要：为探讨化学杂交剂SQ-1诱导的小麦生理型雄性不育系花药绒毡层细胞凋亡过程与花粉粒败育的关系,以小麦生理型雄性不育系及其可育系花药为试验材料,结合前期绒毡层细胞凋亡研究的结果,采用透射电镜超微观察花药绒毡层细胞结构,定量检测与凋亡相关的天冬氨酸蛋白酶相关基因(APs1、APs2、APs3)的表达量,同时利用含明胶的十二烷基硫酸钠聚丙烯酰胺凝胶电泳技术对天冬氨酸蛋白酶活性进行验证。结果表明,在小麦花药发育单核期,生理型雄性不育系花药绒毡层细胞较可育系绒毡层细胞提前降解;在四分体至三核时期,3个天冬氨酸蛋白酶基因在不育和可育系中均表现为先升高后降低的趋势,且单核期表达量最高;天冬氨酸蛋白酶相关基因(APs1、APs2、APs3)在花药发育单核时期以及四分体时期,不育系较可育系均明显上升;天冬氨酸蛋白酶活性研究表明,在花药发育各时期该酶活性在不育系与可育系中均表现出明显差异,且在不育及可育系花药中天冬氨酸蛋白酶活性都呈先升高后降低的趋势,在二核期活性达到最高。表明在SQ-1诱导的生理型不育系中,天冬氨酸蛋白酶的变化与绒毡层细胞凋亡、结构变化及雄性不育花粉粒败育密切相关。本研究初步揭示了生理型小麦败育的机理,为进一步深入研究小麦雄性不育机理,培育能广泛应用于生产实践的优良不育系提供了一定的理论依据。The present study aims at illustrating the relationship between the degradation of tapetum of anther and pollen sterility in physiological male sterility wheat that was induced by SQ-1. The physiologically male sterility and it＇ s corresponding normal lines of complete fertility were used as experimental materials,the ultrastructure of tapetum was analyzed via transmission electron microscope,the APs genes expression were analyzed through Real-time PCR and the activity of aspartic proteinase was analyzed by SDS-Gelatin-Page. Results showed the tapetum of anther was pre-degraded at mononuclear stage in physiological male sterile lines when compared with the normal one. The expressions of 3 APs genes rose firstly and fell from metaphase to three-nuclear stage,and reached their highest expressions at mononuclear stage. Comparing the normal lines,the expressions of APs1,Aps2 and APs3 in physiological male sterile lines showed a higher increase at mononuclear stage and metaphase. The levels of APs activity were firstly upregulated and then decreased,reaching their highest points at binucleate stage. And the activity of APs in physiological male sterile lines was much higher than the normal lines in all stages. These data demonstrated that the higher expression of APs gene in male sterility induced by SQ-1 leads to pre-degradation of tapetum and then causes male sterility.

关 键 词：小麦 化学杂交剂SQ-1 生理型雄性不育 绒毡层 天冬氨酸蛋白酶 

分 类 号：S512.1[农业科学—作物学]