Chitinase,
an important defensive factor in plants, is closely related to plant disease
resistance. In order to understand the expression and function of chitinase
genes, by the using of primer ChiF/ChiR, chitinase genes were cloned by PCR
from 10 wheat cultivars with different resistance to disease in this research.
Source: Baidu
After that, these sequences were analyzed by different biology software, and
were expressed in E. coli. The expression of these genes in root, stem and
leaf were analyzed by quantitative real-time PCR. The highest expression of
chitinase gene in these tissues was selected to test the resistance with fungal
diseases.
The result showed that 5 chitinase gene sequences (960±3 bp in length) were
cloned from 10 wheat cultivars with different resistance to disease. Of them,
one sequence is consistent with the published Chi-3 gene sequence
with the accession number KJ507387, and the others were
named Cht1 to Cht4 with the accession numbers of KR049247
to KR049250, respectively.
Sequence analysis showed that the chitinase genes in this research have no
intron, encoding chitinase belonging to ClassⅠb type. The chitinase
are the members of glycoside hydrolase family 19, which are extracellular
secreted protein and have lysozyme activity.
The results of SDS-PAGE showed that the recombinant plasmid pE-Chi could
express a 33 kDa protein after induction in BL21 (DE3).
The results of quantitative real-time PCR showed that these five kinds of
chitinase genes were expressed in wheat roots, stems and leaves. However, the
amounts of expression in different organs are quite different. The content of
each chitinase gene in this experiment is highest in leaf, followed by root,
and is the minimum in stem.
Meanwhile, the expression of these five kinds of
chitinase genes in the same tissue is ranked
as: Cht4> Cht2> Cht3> Cht1> Chi-3.
Therefore, Cht4 gene, which has the maximum content in the same
organ, was selected to validate its resistance to fungal diseases.
The results showed that under the stripe rust stress, the expression
of Cht4 gene in 92R137 (resistant material) was significantly higher
than in Mingxian 169 (susceptible material), and both of them were higher than
control.
In this research, the chitinase genes are constitutively expressed in
wheat, indicating their participation in the regular growth and development of
wheat. The expression of Cht4 gene is induced by stripe rust,
suggesting that it may be involved in the defense reaction for stripe rust in
wheat leaves.
This article comes from Seed China News 1606, CCM
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Tag: wheat