Cloning and functional analysis of LTP gene in pecan pollen
[Objective]The protein profiles of pecan(Carya illinoinensis(Wangenh.)K.Koch)and Chinese hickory(Carya cathayensis Sarg)were compared before and after pollen germination,screening for significantly different proteins that may play a role in pollen germination.The biological properties and gene expression characteristics of these proteins were then analyzed.[Method]The iTRAQ technology was used to compare the differences of proteome changes of the two Carya plants before and after pollen in vitro germination.The significantly different proteins were identified and their genes were cloned and analyzed.Using the established pecan in vitro germination system and prokaryotic fusion protein expression system,the function of LTP in pollen germination was preliminarily verified.[Result]A comparison of protein ratios showed that some proteins had opposite patterns of change before and after pollen germination in these two plants.This included an 18.6-fold ratio for non-specific lipid-transfer protein.Two gene fragments named CiLTP1 and CiLTP2 were obtained through gene cloning and the CiLTP1 showed better stability in theory.The expression analysis in pecan indicated a high correlation between the LTP homologous genes and pollen germination.After introducing LTP1 into the pollen germination system,it was found that LTP1 does not significantly affect the pollen germination rate and pollen tube length of pecan.[Conclusion]LTP(lipid transfer proteins)were screened out by protein profiles of two species before and after pollen germination.CiLTP1 and CiLTP2 were cloned from pecan and their high correlation with pollen germination was confirmed at the gene expression level.However,the CiLTP1 protein expressed in vitro did not show any influence on the in vitro germination of pecan pollen.This study is an important reference for revealing the pollen germination law of Carya plants.
pecanpollen germination in vitrolipid transfer proteinplant-endophyte bacteria
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维普
