Brill Laurence M, Fujishige Nancy A, Hackworth Cheryl A, Hirsch Ann M
Department of Molecular, Cell and Developmental Biology, Molecular Biology Institute, University of California, 405 Hilgard Avenue, Los Angeles, CA 90095-1606, USA.
Mol Plant Microbe Interact. 2004 Jan;17(1):16-26. doi: 10.1094/MPMI.2004.17.1.16.
Legume lectins have been proposed to have important symbiotic roles during Rhizobium-legume symbioses. To test this hypothesis, the symbiotic responses of transgenic alfalfa plants that express a portion of the putative alfalfa lectin gene MsLEC1 or MsLEC2 in either the antisense or sense orientation were analyzed following inoculation with wild-type Sinorhizobium meliloti 1021. MsLEC1-antisense (LEC1AS) plants were stunted, exhibited hypernodulation, and developed not only abnormally large nodules but also numerous small nodules, both of which senesced prematurely. MsLEC2-antisense plants were intermediate in growth and nodule number compared with LEC1AS and vector control plants. The symbiotic abnormalities of MsLEC1-sense transgene plants were similar to but milder than the responses shown by the LEC1AS plants, whereas MsLEC2-sense transgene plants exhibited symbiotic responses that were identical to those of vector and nontransgenic control plants. MsLEC1 mRNA accumulation was not detected in nodule RNA by Northern blot analysis but was localized to alfalfa nodule meristems and the adjacent cells of the invasion zone by in situ hybridization; transcripts were also detected in root meristems. A similar spatial pattern of MsLEC2 expression was found by using a whole-mount in situ hybridization procedure. Moreover, mRNAs for an orthologous lectin gene (MaLEC) were detected in white sweetclover (Melilotus alba) nodules and root tips.
豆科植物凝集素被认为在根瘤菌与豆科植物的共生过程中具有重要的共生作用。为了验证这一假设,对接种野生型苜蓿中华根瘤菌1021后,以反义或正义方向表达部分假定的苜蓿凝集素基因MsLEC1或MsLEC2的转基因苜蓿植株的共生反应进行了分析。MsLEC1反义(LEC1AS)植株生长受阻,表现出超结瘤现象,不仅形成了异常大的根瘤,还形成了许多小根瘤,且两者均过早衰老。与LEC1AS和载体对照植株相比,MsLEC2反义植株的生长和根瘤数量处于中间水平。MsLEC1正义转基因植株的共生异常与LEC1AS植株表现出的反应相似但较轻,而MsLEC2正义转基因植株表现出与载体和非转基因对照植株相同的共生反应。通过Northern印迹分析未在根瘤RNA中检测到MsLEC1 mRNA积累,但通过原位杂交将其定位到苜蓿根瘤分生组织和侵入区的相邻细胞;在根分生组织中也检测到了转录本。通过整装原位杂交程序发现了MsLEC2表达的类似空间模式。此外,在白花草木樨(Melilotus alba)的根瘤和根尖中检测到了同源凝集素基因(MaLEC)的mRNA。