| A new dwarf and delayed growth mutant caused by T-DNA insertion in rice was identified from 45 000 independent Ds transfer lines. It showed delayed growth, dwarfism, black-green leaf color, large angle of leaf to stem and short root. Genetic analysis indicated that the mutant phenotype was controlled by a single recessive gene. By application the Basta on the healthy leaves of T1 and F2 generations showed that the mutant was co-segregated with Basta resistance. PCR and Southern blotting further confirmed the above analysis.OsWRKY 10 gene, which showed high identity with PAC clones of AP002744 and AP002839, located in chromosome 1 of rice, was amplified from DNA fragment flanking of T-DNA insertion mutant by using thermal asymmetric interlaced PCR(Tail-PCR). DNA and protein sequence alignment revealed that the genome DNA of OsWRKY10 is 2272 bp in length, encodes a predicated polypeptide of 381 amino acids and an open reading fragment of 1118 bp, the deduced protein harbors one WRKY conserved domain and a C2H2 zinc finger motif, is classified into the group 2a of OsWRKY transcriptional factor family. Additionally, it contains a nuclear localization signal (NLS) and rich regulation protein motif.Plant WRKY genes encoded transcriptional regulators appear to be involved invarious physiological programs, including biotic and abiotic stress responses as wellas development processes. To study the functions of the OsWRKY10 and othermembers in OsWRKY superfamily in rice, we constructed a hairpin dsRNAinterference vector involving in a sequence of WRKY conserved domain, constitutedby about 60 amino acid residues, and was transformed into the embryos of wild parentrice Zhonghua 11 by Agrobacterium-mediated co-cultivation method. Nine transgeniclines with interference phenotypes were identified from 65 independent transgenicplants, so the interference rate was 13.8%. PCR and Southern blotting analysisindicated that foreign DNA had been integrated into rice genome DNA of Zhonghua11 , and most of them were single copy. The resistance to rice blast (Magnaporthegrisea) and rice bacterial blight (Xanthomonas oryzae PV. oryzae) was conducted onT-DNA mutant (named T456), Zhonghua 11 and the three interference lines, the... |
