甘蓝型油菜新型细胞质雄性不育类型NRO4270A的鉴定
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国家高技术研究发展计划(2012AA101107);国家科技支撑计划(2011BAD35B04);公益性行业科研专项(201203026-6)


Identification of a New Cytoplasmic Male Sterility Type NRO4270A in Brassica napus
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    摘要:

    甘蓝型油菜细胞质雄性不育系(CMS)NRO4270A是从萝卜甘蓝(Raphanus brassica,RRCC)与甘蓝型油菜(Brassica napus,AACC)远缘杂交后代中发现的一种CMS类型,该研究通过育性调查、恢保关系测定、花药发育的显微观察和线粒体DNA的RFLP分析对NRO4270A不育系进行鉴定。结果表明:(1) 通过连续5年在冬油菜区和春油菜区的育性调查,发现NRO4270A不能产生花粉,败育彻底,不育性稳定,不受温度、光照等环境条件的影响,不育率和不育度均为100%。(2) 将NRO4270A、pol、ogu、kos、hau等CMS分别与pol、ogu恢复系以及‘华双4号’测交,显示NRO4270A与pol、ogu、kos等不育系统的恢保关系明显不同。(3) 通过对NRO4270A与保持系NRO4270B花药发育的显微结构观察和比较,表明NRO4270A花粉的败育时期为四分体期至单核花粉晚期,败育特点为:单核小孢子时期,单核花粉粒不能形成外壁,花药绒毡层液泡化,并逐渐膨大增厚,最后绒毡层和小孢子彻底消失,花粉囊变为空腔,花药完全败育。(4) 利用18个探针/酶组合(由6个线粒体基因atp1、atp6、atp9、orf222、cox2、cob探针和3种限制性内切酶EcoRⅠ、BamHⅠ和HindⅢ组成)对NRO4270A、pol、ogu、kos、hau等CMS以及NRO4270B和‘华双5号’的线粒体DNA进行RFLP分析,在11个探针/酶组合中,NRO4270A均显示与其他材料明显不同的带型,表明其为新型的不育胞质类型。NRO4270A的发现和利用将有利于克服中国油菜主要CMS类型对环境的敏感性和生产上不育胞质类型单一化等问题。

    Abstract:

    Brassica napus cytoplasmic male sterility (CMS) NRO4270A was obtained in the progenies of distant hybridization of Raphanus brassica (RRCC) and B.napus (AACC).(1)The fertility investigations of five consecutive years in both winter and spring rapeseed production areas showed that NRO4270A could not produce pollens completely and its male sterility was extremely stable,which could not be affected by environmental conditions,such as temperature and photo period.Its sterility rate and sterility degree are both 100%.(2)NRO4270A,pol,ogu,kos,hau CMS lines were test-crossing with the restorer lines of pol,ogu as well as cultivar Huashuang 4.It was suggested that NRO4270A had different restorer and maintainer relationship with pol,ogu and kos.(3)Microscopic structure analysis and comparison of anther developments of NRO4270A and its maintainer line NRO4270B indicated that NRO4270A pollen abortions occurred between the tetrad stage and the mononucleate stage.At mononucleate stage,its microspore exine could not form,and its tapetum was vacuolated and expanded.Eventually,its tapetum and microspore were completely degraded.The pollen sacs became empty and the pollens could not be produced in it.(4)The mitochondrial DNAs of NRO4270A,pol,ogu,kos,hau,NRO4270B and Huashuang 5 were analyzed through RFLP by 18 probe/enzyme combinations (six mitochondrial gene,atp1,atp6,atp9,orf222,cox2 and cob,as well as three restriction enzymes,EcoRⅠ,BamHⅠ and HindⅢ),respectively.In 11 combinations,NRO4270A showed significantly different bands with the other materials,suggesting that NRO4270A was a new CMS type.Its discovery and utilization will overcome the problems of environmental sensitivity of Chinese major CMS system and the singleness of sterile cytoplasm type in rapeseed hybrid seed production.

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舒 畅,刘 超,吴江生.甘蓝型油菜新型细胞质雄性不育类型NRO4270A的鉴定[J].西北植物学报,2015,35(5):898-905

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  • 在线发布日期: 2015-06-09
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