Type Ⅲ secretion system 1 BPSS1395 gene promotes
Burkholderia pseudomallei colonization and proliferation in tomato seedlings- Journal of Third Military Medical University Vol. 42, Issue 10, Pages: 973-980(2020)
- 作者机构：
1.400715 重庆，西南大学生命科学学院，三峡地区生态环境与生物资源国家重点实验室培育基地，现代生物制药研究所
2.400038 重庆，陆军军医大学药学与检验医学系临床微生物与免疫学教研室
- 作者简介：
- 基金信息：the General Program of National Natural Science Foundation of China(81772141);the General Program of National Natural Science Foundation of China(81971907)
DOI：10.16016/j.1000-5404.202001062
CLC： Q933;Q949.777.7;R378.99Received：08 January 2020，
Revised：2020-03-06，
Published：30 May 2020
- 稿件说明：
移动端阅览
Siqi YUAN, Yi HU, Mengling TANG, et al. Type Ⅲ secretion system 1 BPSS1395 gene promotes
Burkholderia pseudomallei colonization and proliferation in tomato seedlings[J]. Journal of Third Military Medical University, 2020, 42(10): 973-980.Siqi YUAN, Yi HU, Mengling TANG, et al. Type Ⅲ secretion system 1 BPSS1395 gene promotes
Burkholderia pseudomallei colonization and proliferation in tomato seedlings[J]. Journal of Third Military Medical University, 2020, 42(10): 973-980. DOI： 10.16016/j.1000-5404.202001062.
摘要
目的
2
研究类鼻疽菌T3SS1的BPSS1395基因在番茄植株感染中的作用。
方法
2
类鼻疽菌野生株BPC006感染番茄幼苗根部，以大肠杆菌感染组作为对照，通过PCR技术检测番茄幼苗叶片中细菌的存在，并通过透射电镜观察番茄幼苗叶片细胞中类鼻疽菌的定位。类鼻疽菌感染第3、7、10、14、17、21天后，随机选取幼苗叶片，通过胞内存活实验检测类鼻疽菌在叶片细胞中的增殖。采用同源重组技术构建BPSS1395基因的敲除株和回补株，分别以类鼻疽菌野生株BPC006、敲除株Δ
BPSS 1395
、回补株c-Δ
BPSS 1395
感染番茄幼苗根部后，首先直接观察叶片的病理变化；其次提取感染第14和21天各组植物叶片的基因组，通过PCR对细菌的定植进行鉴定；最后采用胞内存活实验检测各菌株在叶片细胞内的增殖情况。
结果
2
类鼻疽菌感染番茄植株后，在提取的番茄幼苗叶片基因组中，能扩增出BPSS1395基因条带，而对照组未扩增出大肠杆菌特异性条带；通过透射电镜观察到类鼻疽菌存在于叶片细胞中，且叶片细胞内的类鼻疽菌数量随着感染时间的延长而增加。敲除株Δ
BPSS 1395
感染番茄幼苗后，叶片无任何疾病症状，而野生株BPC006和回补株c-Δ
BPSS 1395
感染组的叶片有些许的萎蔫。此外，在敲除株Δ
BPSS 1395
感染组提取的植物基因组中未检测到类鼻疽菌的存在，而回补株c-Δ
BPSS 1395
表现出和野生株相似的叶片细胞内的增殖能力。
结论
2
类鼻疽菌T3SS1的BPSS1395基因有利于该菌在番茄幼苗感染中的定植和增殖。
Abstract
Objective
2
To investigate the role of type Ⅲ secretion system 1 BPSS1395 gene of
Burkholderia pseudomallei (B. pseudomallei)
in promoting the infection of the bacterium in tomato seedlings.
Methods
2
Using
Escherichia coli(E.coli)
as the control
we infected the roots of tomato seedlings with wide-type strain
B. pseudomallei
BPC006 and detected the presence of the bacteria in the leaves of the plant with PCR. The localization of
B. pseudomallei
in tomato leaves was observed with transmission electron microscopy. At 3
7
10
14
17 and 21 d after the infection
the leaves of the tomato seedlings were randomly selected for detecting the intracellular survival of
B. pseudomallei
. We also compared the results of infections of the tomato seedlings with the wide-type
B. pseudomallei
strain BPC006
the Δ
BPSS 1395
mutant strain and the BPSS1395-complemented strain by observing the pathological changes and PCR detection of bacterial colonization in the leaves on days 14 and 21 following the infection; the proliferation of each strain in the cells of the leaves was examined by counting the intracellular bacteria.
Results
2
A distinct BPSS1395 gene band could be amplified from the leaves of the tomato seedlings in
B. pseudomallei
infected group
while no
E. coli-specific
band was observed in the control group. Transmission electron microscopy revealed the presence of
B. pseudomallei
in the leaves after infection of the roots of the tomato seedlings
and the number of
B. pseudomallei
increased over time following the infection. Infection with Δ
BPSS 1395
mutant resulted in no obvious pathologies in the leaves of the tomato seedlings
while the leaves of the seedlings infected with the wild-type strain BPC006 and BPSS13
95-complemented strain were slightly wilted. Furthermore
no
B. pseudomallei
was detected in the leaves of the seedlings following infection with the Δ
BPSS 1395
mutant; the BPSS1395-complemented strain and the wild-type strain showed similar proliferation ability in the leaf cells.
Conclusion
2
BPSS1395 gene can promote the colonization and proliferation of
B. pseudomallei
in tomato seedlings.
关键词
Keywords
references
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