棉花GhHB基因克隆和表达初步分析-生化与分子生物学专业毕业论文.docxVIP

A．bstractHomeodomain A．bstract Homeodomain leucine zipper(HD-Zip)is one type of transcriptional factor．Its structuresconsistofahomeodomain and alencinezipperregion．HD-Zipencodinggenes ale identified only in plant．HD-Zip plays all important role in morphological changes of plant，such as leaf development,stem elongtion，hormone response and so on．In our study,we isolate four HD-Zip encoding genes．Structure,conservation and function four genes are investigated．n地results as follows： 1．Wre isolate four HD—Zip protein-encoding genes．named GhHBI(Gossypium hirsutum homeobox)I．GhHB2,GhHB3 and GhHB4．The elementary anlysis of four HD-Zip protein sequences from couon(Gossypium hirsutum)show that all ofthem have many PKC PHOSPHO SrrEs and CK2 PHOSPH0 SnEs．The analysis also shows that they possess homedomain and lencine zipper region． 2．Phylogenctie analysis indicates that all 44 HD—Zip proteins from different plant ale divided into two clades．GhHBl．G1羽[B2，GhHB3 and GllHB4 ale belonged to those two clades，suggesting that those genes may have very close evolutionary relationship． Gh衄1，G11HB2，GhHB3 and Gll}IB4 have the closest relationship with OsHox3，HAT4， P#m3 and ANL2，respectly． 3．RNA was extracted from coRon tissues；we analyzed the expression patterns of GhHBl．GhHB2,GhHB3 and GhHB4 by realtime SYBR-Green fluorescence quantity P瓯The results indicate that Ghlibj．G^瑚2J G^瑚3 and G^册4 are predominantly expressed in roots and hypocotyls，suggesting that it may function in roots and hypocotyls development．GhHB4 is also expressed in eotyledon．GhHBl expression in roots and hypocotyls is also up—regulated by NaCL GA3 and ABA，suggestting that the GhHBl may play all important role in response to salt-stresses and to different phytohormone sigualings， 4．Different GhHB3 fragment was cloned into pPZP221 and pBIl21 to obtain overexpression vector end supression vector respectively．GhHB3 fragment was transformed into cotton by Agrobacterium-mediated transformation method．A great number of transfo