qPCR analysis and RNAi define pharyngeal gland cell-expressed genes of Heterodera glycines required for initial interactions with the host
qPCR analysis and RNAi define pharyngeal gland cell-expressed genes of Heterodera glycines required for initial interactions with the host
复制标题
DOI：
10.1094/mpmi-20-3-0306
复制
发表时间：
2007-03-01
影响因子：
3.5
通讯作者：
Atkinson, H. J.
中科院分区：
文献类型：
--
作者：
Bakhetia, M.;Urwin, P. E.;Atkinson, H. J.
通过定量聚合酶链反应（qPCR）监测宿主入侵后大豆胞囊线虫（Heterodera glycines）的三种咽腺细胞中表达的基因的转录丰度的变化，并在入侵前通过RNAi处理研究破坏其表达的后果。已知两个转录本在两个腹下腺细胞中表达（hg-pel和hg-eng-1），另外两个仅在单个背腺细胞中表达（hg-gp和hg-syv 46），第五个转录本（hg-cm）由两种腺细胞类型表达。qPCR研究证实，hg-syv 46和hg-gp的转录物在感染后2天（dpi）大量增加，前者保持最丰富。从0到14 dpi，hg-cm转录水平显示出微小的变化，但在21 dpi时确实下降。相比之下，hg-eng-1和hg-eng-2信使（m）RNA下降7 dpi和hg-pel的14 dpi，然后在21 dpi增加。hg-eng-1的RNAi靶向减少了第10天植物上存在的雌性数量。针对汞糖蛋白，汞厘米，汞pel引起的性别命运的变化，有利于男性发展的根。在靶向hg-syv 46后，这两种效应都很明显。RNAi对第二阶段幼鱼（J2（i））hg-eng-1 mRNA水平的抑制是短暂的，处理后在水中孵育15天后恢复。预浸H.具有双链RNA的甘氨酸J2对研究线虫与植物互作早期的基因功能具有重要价值。
Changes in transcript abundance of genes expressed in the three pharyngeal gland cells of Heterodera glycines after host invasion were monitored by quantitative polymerase chain reaction (qPCR) and the consequences of disrupting their expression studied by RNAi treatment prior to invasion. Two transcripts were known to be expressed in the two subventral gland cells (hg-pel and hg-eng-1), a further two in the single dorsal gland cell only (hg-gp and hg-syv46), and a fifth transcript (hg-cm) was expressed by both gland cell types. The qPCR study established that transcripts of hg-syv46 and hg-gp increased in abundance by 2 days postinfection (dpi), with the former remaining the most abundant. The hg-cm transcript level showed minor changes from 0 to 14 dpi but did fall by 21 dpi. In contrast, hg-eng-1 and hg-eng-2 messenger (m)RNA declined by 7 dpi and hg-pel by 14 dpi before it increased at 21 dpi. RNAi-targeting of hg-eng-1 reduced the number of females present on the plants at 10 days. Targeting of hg-gp, hg-cm, and hg-pel caused a change in sexual fate favoring male development on roots. Both effects were evident after targeting hg-syv46. Suppression of hg-eng-1 mRNA levels in second-stage juveniles (J2(i)) by RNAi was transient, with a recovery by 15 days of incubation in water after treatment. Presoaking H. glycines J2 with double-stranded RNA has value for studying gene function during the nematode's early interaction with a plant.