Cloning and characterization of a balsam pear class I chitinase gene (Mcchit1) and its ectopic expression enhances fungal resistance in transgenic plants.
A balsam pear (Momordica charantia L.) chitinase (Mcchit1) was purified and sequenced at the N-terminal. The genomic and cDNA coding sequences of Mcchit1 were cloned by rapid amplification of 3' cDNA ends (3'-RACE) and the Y-shaped adaptor dependent extension (YADE) method. Sequence analysis showed that the Mcchit1 protein is a class I chitinase containing a chitin-binding domain and a catalytic domain, but no C-terminal extension. Northern blot indicated that the Mcchit1 transcription is wound-inducible. Overexpression of Mcchit1 dramatically increased intercellular and intracellular endochitinase activities, suggesting that the Mcchit1 gene encodes a secretory endochitinase. It was also found that overexpression of Mcchit1 significantly enhanced resistance to the plant pathogenic fungus Phytophthora nicotianae in transgenic N. benthamiana plants and against Verticillium wilt in transgenic cottons, indicating that the Mcchit1 gene can be a useful gene in plant engineering against fungal diseases.
Xiao
Yue-Hua
YH
Key Laboratory of Biotechnology and Crop Quality Improvement of the Ministry of Agriculture of China, Biotechnology Research Center, Southwest University.
Li
Xian-Bi
XB
Yang
Xing-Yong
XY
Luo
Ming
M
Hou
Lei
L
Guo
Shao-Hui
SH
Luo
Xiao-Ying
XY
Pei
Yan
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D020134:Catalytic Domain; D002688:Chitinases / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005805:Genes, MHC Class I; D017343:Genes, Plant; D008969:Molecular Sequence Data; D029777:Momordica charantia / Q000201:enzymology / Q000235:genetics* / Q000382:microbiology; D010838:Phytophthora / Q000502:physiology*; D010935:Plant Diseases / Q000382:microbiology; D010940:Plant Proteins; D030821:Plants, Genetically Modified; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D020172:Verticillium / Q000502:physiology*
D018076:DNA, Complementary; D018744:DNA, Plant; D010940:Plant Proteins; D002688:Chitinases; C119825:chitinase C