Cloning of phospholipase D from grape berry and its expression under heat acclimation.
To investigate whether phospholipase D (PLD, EC 3.1.4.4) plays a role in adaptive response of post-harvest fruit to environment, a PLD gene was firstly cloned from grape berry (Vitis Vinifera L. cv. Chardonnay) using RT-PCR and 3'- and 5'-RACE. The deduced amino acid sequence (809 residues) showed 84.7% identity with that of PLD from Ricinus communis. The secondary structures of this protein showed the characteristic C2 domain and two active sites of a phospholipid-metabolizing enzyme. The PLD activity and its expression in response to heat acclimation were then assayed. The results indicated PLD was significantly activated at enzyme activity, as well as accumulation of PLD mRNA and synthesis of new PLD protein during the early of heat acclimation, primary suggesting that the grape berry PLD may be involved in the heat response in post-harvest grape berry. This work offers an important basis for further investigating the mechanism of post-harvest fruit adaptation to environmental stresses.
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
Si-Bao
SB
Wan
Wei
W
Wang
Peng-Fei
PF
Wen
Jian-Ye
JY
Chen
Wei-Fu
WF
Kong
Qiu-Hong
QH
Pan
Ji-Cheng
JC
Zhan
Li
L
Tian
Hong-Tao
HT
Liu
Wei-Dong
WD
Huang
D000064:Acclimatization / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000378:metabolism; D005638:Fruit / Q000201:enzymology* / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant / Q000235:genetics; D006360:Heat-Shock Proteins / Q000378:metabolism; D006358:Hot Temperature; D008969:Molecular Sequence Data; D010739:Phospholipase D / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D027843:Vitis / Q000201:enzymology* / Q000235:genetics
D018076:DNA, Complementary; D018744:DNA, Plant; D006360:Heat-Shock Proteins; D010739:Phospholipase D