Lohar Dasharath, Stiller Jiri, Kam Jason, Stacey Gary, Gresshoff Peter M
ARC Centre of Excellence for Integrative Legume Research, The University of Queensland, St Lucia, Brisbane, QLD 4072, Australia.
Ann Bot. 2009 Aug;104(2):277-85. doi: 10.1093/aob/mcp132. Epub 2009 Jun 7.
Transgenics are used to demonstrate a causal relationship between ethylene insensitivity of a seedling legume plant, the level of ethylene receptor gene expression, lateral root growth and Mesorhizobium loti-induced nodule initiation.
Lotus japonicus plants expressing the dominant etr1-1 allele of the Arabidopsis thaliana gene encoding a well-characterized mutated ethylene receptor were created by stable Agrobacterium tumefaciens transformation. Single insertion, homozygous lines were characterized for symbiotic properties.
Transgenic plants were ethylene insensitive as judged by the lack of the 'Triple Response', and their continued ability to grow and nodulate in the presence of inhibitory concentrations of ACC (1-aminocyclopropane-1-carboxylic acid; an ethylene precursor). Transgenic plants with high insensitivity to ACC had significantly fewer lateral roots and exhibited increased nodulation while showing no altered nitrate sensitivity or lack of systemic autoregulation. Whereas ACC-insensitive shoot growth and nodulation were observed in transformants, root growth was inhibited similarly to the wild type. Increased nodulation was caused by increased infection and a seven-fold increase in nodules developing between xylem poles. Bacteroid numbers per symbiosome increased about 1.7-fold in ethylene-insensitive plants.
The study further demonstrates multiple roles for ethylene in nodule initiation by influencing root cell infections and radial positioning, independent of autoregulation and nitrate inhibition of nodulation.
利用转基因技术来证明豆科幼苗植物的乙烯不敏感性、乙烯受体基因表达水平、侧根生长以及百脉根诱导的根瘤起始之间的因果关系。
通过根癌农杆菌稳定转化,培育出表达拟南芥基因etr1-1显性等位基因的日本百脉根植株,该基因编码一种特征明确的突变乙烯受体。对单插入纯合系的共生特性进行了表征。
转基因植株表现出乙烯不敏感性,这可通过缺乏“三重反应”来判断,并且在存在抑制浓度的ACC(1-氨基环丙烷-1-羧酸;一种乙烯前体)的情况下,它们仍具有生长和形成根瘤的能力。对ACC高度不敏感的转基因植株侧根明显较少,根瘤形成增加,同时硝酸盐敏感性未改变,也不存在系统性自调控缺失的情况。虽然在转化体中观察到对ACC不敏感的地上部生长和根瘤形成,但根的生长与野生型类似受到抑制。根瘤形成增加是由于感染增加以及木质部极之间形成的根瘤增加了7倍。乙烯不敏感植株中每个共生体中的类菌体数量增加了约1.7倍。
该研究进一步证明了乙烯通过影响根细胞感染和径向定位在根瘤起始中发挥多种作用,这与自调控以及硝酸盐对根瘤形成的抑制无关。