K型温敏雄性不育小麦KTM3315A的鉴定及花粉败育特点的初步分析

Identification and characteristics of KTM3315A,a thermo-sensitive male sterile line with Aegilops kotschyi cytoplasm

为了解小麦温敏雄性不育系KTM3315A的染色体组成,揭示其雄性败育特点,采用分子标记和A-PAGE技术对KTM3315A是否含有T1BL.1RS易位染色体进行了鉴定。通过醋酸洋红、DAPI和I2-KI染色,观察花粉败育的细胞学特点及类型,并对不同发育时期(减数分裂、单核早期、单核晚期、二核期和三核期)花药中的保护酶(POD、SOD和CAT)活性进行了测定。结果表明:KTM3315A具有小麦1BS的特异扩增条带,但缺少黑麦1RS的特异扩增条带,与KTM3315A在ω区不具有黑麦特异蛋白谱带的结果相对应,推测KTM3315A为非1B/1R类型的K型温敏雄性不育小麦;KTM3315A在不育条件下的三核期花粉粒经I2-KI染色表现为染色不均匀,花粉败育,其花粉败育类型为染败;3种活性氧代谢的保护酶活性在不同育性条件下呈现出不同的变化趋势,在不育和可育条件下,POD、SOD和CAT活性在单核晚期均表现显著差异,推测KTM3315A的雄性不育与活性氧代谢的保护酶活性有关,其败育发生的关键时期可能在单核晚期。

To understand the chromosome constitution of thermo-sensitive male sterile wheat line KTM3315 Aand to its male sterility characteristics,T1 BL.1RS translocation chromosome was identified by molecular markers and A-PAGE.The cytological characteristics and type of pollen abortion were observed by staining of pollen grains by aceto carmine,DAPI and I2-KI.The activities of protective enzymes（POD,SOD,CAT）in anther of KTM3315 A were determined at different development stages（tetrad stage,early uninucleate stage,late uninucleate stage,binucleate stage,trinucleate stage）.Molecular marker electrophoresis results showed that KTM3315 Acould be amplified bands in the marker of wheat chromosome 1BS,but not amplified bands in the marker of chromosome 1RS in rye.KTM3315 Adid not have the same bands inωregion as rye in A-PAGE electrophoresis.Therefore,KTM3315 A was non-1B/1R K type thermosensitive male sterile wheat line.Its pollens was not fully dyed by I2-KI at trinucleate stage under sterile conditions,which proved pollens were abortive and the type of pollen abortion was stainable abortion.The activity of their protective enzymes was different in both sterile and fertile conditions.POD,SOD and CAT activity showed significant differences at late uninucleate stage,suggesting that male sterility was associated with activities of protective enzymes in active oxygen metabolism.KTM3315 Aabortion mainly occurred at late uninucleate stage.