Cloning And Expression Analysis Of The Melon’s CmMAPK、CmBADH、CmP5CS Genes And Transformation

Melon(Cucumis melo L.)is one of the most important horticultural crops in the world,Soil secondary salinization and extreme temperature is one of the major limited factorsaffecting its production in the greenhouse. The clone of these genes and their expressions aresignificant for revealing the stress tolerance mechanmism at molecular level and improvingthe stress resistance of melon. In this study, the full-length cDNA sequence of the key prolinesynthetase gene CmP5CS, the full-length cDNA sequence of the key betaine aldehydedehydrogenase synthetase gene CmBADH, and a full-length cDNA sequence of themitogen-activated protein kinase(CmMAPK), which transfer stress signals, were cloned. Wealso analyzed these genes expression characteristic and their genetic transformation to melon.The main results are as follows:(1) We cloned three genes(CmMAPK、CmBADH、CmP5CS) from muskmelon TS-2byRT-PCR and3’,5’RACE. The full length sequence of CmBADH was1856bp, encoding503amino acids and the registration number in GenBank is JN091961, and the full lengthsequence of CmMAPK was1391bp, encoding371amino acids and the registration number inGenBank is JF820842, and the full length sequence of CmP5CS was2761bp, encoding718amino acids and the registration number in GenBank is JN590050.(2) qPCR analysis showed the expression of CmP5CS, CmBADH is induced by salt,drought, low and high temperature as well as ABA treatment; the expression of CmMAPK isinduced by salt, drought and low temperature treatment but not induced by high temperaturetreatment(3) Sulfosalicylic acid method analysis indicated that proline content were increased indifferent level under salt, drought, low temperature, high temperature and ABA treatment.(4) antisense expression vector CmP5CS, CmMAPK were constructedAgrobacterium-mediated method was used to genetic transformed antisense expression vectorinto the melon, respectively.