Interspecies compatibility of the anther specific cell wall invertase promoters from Arabidopsis and tobacco for generating male sterile plants
Interspecies compatibility of the anther specific cell wall invertase promoters from Arabidopsis and tobacco for generating male sterile plants
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DOI：
10.1007/s00122-008-0892-2
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发表时间：
2009-01-01
影响因子：
5.4
通讯作者：
Roitsch, T.
中科院分区：
文献类型：
--
作者：
Hirsche, J.;Engelke, T.;Roitsch, T.
组织化学GUS染色和荧光分析表明，烟草的细胞壁转化酶启动子NIN88和拟南芥的ATcwINV2分别与花药和花粉紧密结合，具有较强的组织特异性。在拟南芥和烟草中，通过花药特异性RNA干扰抑制转化酶活性是避免共生花粉碳水化合物供应的一种有效方法，从而使花粉萌发能力和结实率显著下降。在烟草中，通过表达一种蛋白类转化酶抑制剂也可以获得类似的结果，而这种方法在拟南芥中的效率较低。本研究表明，花药特异性干扰转化酶活性以产生雄性不育植株可应用于茄科和十字花科两个不同植物家族的成员，这一策略似乎是在杂交育种中实际应用或作为生物安全预防措施的通用工具。为了阐明分离的启动子在植物家族以外的亲和性，我们将调控序列分别转移到了各自的异源系统中，即NIN88启动子进入拟南芥，AtcwINV2启动子进入烟草。两个启动子的特异性在异源背景中保持不变，但随着花和花粉的GUS染色和荧光定量结果的证实，它们的活性显著降低。
Histochemical GUS-staining and fluorometric analyses revealed strong tissue specific activities of the cell wall invertase promoters Nin88 from Nicotiana tabacum and AtcwINV2 from Arabidopsis thaliana that are restricted tightly to anthers and pollen, respectively. Both in A. thaliana and N. tabacum repression of invertase activity by anther specific RNA-interference turned out to be an efficient method to circumvent carbohydrate supply of the symplastically isolated pollen with subsequent strong decrease of pollen germination ability and seed setting. In the case of tobacco, comparable results were also obtained by expressing a proteinaceous invertase inhibitor, whereas this approach was less efficient in Arabidopis. The present study revealed that anther specific interference with invertase-activity in order to generate male sterile plants can be applied to members of the two different plant families Solanaceae ( N. tabacum) and Brassicaceae ( A. thalaina) and the strategy seems to be a general tool for practical application in hybrid breeding or as biological safety precautions. To elucidate the compatibility of the isolated promoters beyond plant families, we transferred the regulatory sequences into the respectively heterologous systems, i. e. the Nin88 promoter into Arabidopsis and the AtcwINV2 promoter into tobacco. The specific of both promoters are maintained in the heterologous backgrounds, but their activities are strongly reduced as GUS-stainings of flowers and pollen revealed and fluorometrical quantification conformed.