Zhang Xueying, Wang Liman, Xu Xiaoyang, Cai Caiping, Guo Wangzhen
State Key Laboratory of Crop Genetics & Germplasm Enhancement, Hybrid Cotton R & D Engineering Research Center, MOE, Nanjing Agricultural University, Nanjing, 210095, Jiangsu Province, P. R. China.
BMC Plant Biol. 2014 Dec 10;14:345. doi: 10.1186/s12870-014-0345-9.
Mitogen-activated protein kinase (MAPK) cascades play a crucial role in plant growth and development as well as biotic and abiotic stress responses. Knowledge about the MAPK gene family in cotton is limited, and systematic investigation of MAPK family proteins has not been reported.
By performing a bioinformatics homology search, we identified 28 putative MAPK genes in the Gossypium raimondii genome. These MAPK members were anchored onto 11 chromosomes in G. raimondii, with uneven distribution. Phylogenetic analysis showed that the MAPK candidates could be classified into the four known A, B, C and D groups, with more MAPKs containing the TEY phosphorylation site (18 members) than the TDY motif (10 members). Furthermore, 21 cDNA sequences of MAPKs with complete open reading frames (ORFs) were identified in G. hirsutum via PCR-based approaches, including 13 novel MAPKs and eight with homologs reported previously in tetraploid cotton. The expression patterns of 23 MAPK genes reveal their important roles in diverse functions in cotton, in both various developmental stages of vegetative and reproductive growth and in the stress response. Using a reverse genetics approach based on tobacco rattle virus-induced gene silencing (TRV-VIGS), we further verified that MPK9, MPK13 and MPK25 confer resistance to defoliating isolates of Verticillium dahliae in cotton. Silencing of MPK9, MPK13 and MPK25 can significantly enhance cotton susceptibility to this pathogen.
This study presents a comprehensive identification of 28 mitogen-activated protein kinase genes in G. raimondii. Their phylogenetic relationships, transcript expression patterns and responses to various stressors were verified. This study provides the first systematic analysis of MAPKs in cotton, improving our understanding of defense responses in general and laying the foundation for future crop improvement using MAPKs.
丝裂原活化蛋白激酶(MAPK)级联在植物生长发育以及生物和非生物胁迫响应中起着至关重要的作用。关于棉花中MAPK基因家族的知识有限,尚未见对MAPK家族蛋白的系统研究报道。
通过进行生物信息学同源性搜索,我们在雷蒙德氏棉基因组中鉴定出28个假定的MAPK基因。这些MAPK成员定位在雷蒙德氏棉的11条染色体上,分布不均。系统发育分析表明,MAPK候选基因可分为已知的A、B、C和D四组,含TEY磷酸化位点的MAPK(18个成员)比含TDY基序的MAPK(10个成员)更多。此外,通过基于PCR的方法在陆地棉中鉴定出21个具有完整开放阅读框(ORF)的MAPK cDNA序列,包括13个新的MAPK和8个先前在四倍体棉花中报道过的同源物。23个MAPK基因的表达模式揭示了它们在棉花营养生长和生殖生长的各个发育阶段以及胁迫响应的多种功能中发挥的重要作用。使用基于烟草脆裂病毒诱导基因沉默(TRV-VIGS)的反向遗传学方法,我们进一步验证了MPK9、MPK13和MPK25赋予棉花对大丽轮枝菌落叶分离株的抗性。沉默MPK9、MPK13和MPK25可显著增强棉花对该病原菌的易感性。
本研究全面鉴定了雷蒙德氏棉中的28个丝裂原活化蛋白激酶基因。验证了它们的系统发育关系、转录表达模式以及对各种胁迫的响应。本研究首次对棉花中的MAPK进行了系统分析,增进了我们对一般防御反应的理解,并为未来利用MAPK改良作物奠定了基础。