Development of wheat scab symptoms is delayed in transgenic wheat plants that constitutively express a rice thaumatin-like protein gene
Development of wheat scab symptoms is delayed in transgenic wheat plants that constitutively express a rice thaumatin-like protein gene
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DOI：
10.1007/s001220051294
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发表时间：
1999-09-01
影响因子：
5.4
通讯作者：
Gill, BS
中科院分区：
文献类型：
--
作者：
Chen, WP;Chen, PD;Gill, BS
小麦赤霉病(由Fusum graminearum Schw.)通过对小麦植株进行工程改造，探索病程相关(PR)蛋白基因的结构性表达。通过共转化pGL2ubi-TLP(泛素/TLP//CaMV 35S/HPT)和pAHG11(CaMV 35S/chi11//ubiquitin/bar)，将水稻thaumatin-like蛋白(TLP)基因和水稻几丁质酶基因(Chill)导入春小麦品种‘BobWhite’。这一转变是通过生物轰炸实现的。以Bialaphos为选择试剂。通过Southern、Northern和Western印迹分析了TIP、bar、chill和hpt基因的整合和表达。以pAHG11或pGL2ubi-TLP为探针，通过连续N显带和荧光原位杂交(FISH)，将4个转基因基因共分离于转基因植株T-1代，并定位于6A染色体长臂的端粒区。只有在泛素启动子-内含子控制下的转基因TIP和BAR被表达。在T-1植株中未检测到由CaMV 35S启动子控制的chill和hpt基因的表达。接种小麦赤霉菌分生孢子后，表达TIP基因的T-1、T-2和T-3代转基因植株的赤霉病症状发展明显慢于未转化的对照植株。这是首次报道了含有TLP基因的转基因小麦植株对禾谷镰刀菌的抗性增强。
The possibility of controlling wheat scab (caused by Fusarium graminearum Schw.) was explored by engineering wheat plants for constitutive expression of pathogenesis-related (PR) protein genes. A rice thaumatin-like protein (TLP) gene (tlp) and a rice chitinase gene (chill) were introduced into the spring wheat cultivar 'Bobwhite' by co-transformation of the plasmids pGL2ubi-tlp (ubiquitin/tlp//CaMV 35S/hpt) and pAHG11 (CaMV 35S/chi11//ubiquitin/bar). The transformation was by biolistic bombardment. Bialaphos was used as the selection reagent. The integration and expression of the tip, bar, chill and hpt genes were analyzed by Southern, Northern and Western blot analyses. The four transgenes co-segregated in the T-1 progeny of the transgenic plant and were localized at the telomeric region of the chromosome 6A long arm by sequential N-banding and fluorescent in situ hybridization (FISH) using pAHG11 or pGL2ubi-tlp as the probes. Only the transgenes tip and bar, under the control of the ubiquitin promoter-intron, were expressed. No expression of the chill and hpt genes, controlled by the CaMV 35S promoter, was detected in T-1 plants. After inoculation with conidia of F. graminearum, the symptoms of scab developed significantly slower in transgenic plants of the T-1, T-2 and T-3 generations expressing the tip gene than in non-transformed control plants. This is the first report of enhanced resistance to F. graminearum in transgenic wheat plants with constitutive expression of TLP.