番茄内生细菌种群和数量动态分析及对番茄青枯病菌的拮抗作用研究
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摘要
从广西武鸣、田阳、桂林、玉林采集270份田间健康的番茄植株进行内生细菌的分离，结果分离到571个细菌菌株，分属于Bacillus Spp．，Pseudomonas spp．，Xanthomonas spp．，Microbacterium spp．，Agrobacterium spp．，Enterobacter spp．，Erwinia spp．和Corynebacterium spp．。其中以Bacillus spp．为优势种群。
番茄内生细菌的种群及数量随植株不同器官、生育期、地区和品种而变化。测试的红宝石品种中，根无Corynebacterium spp．，茎中无Microbacterium spp．，但在美星品种中各器官的内生细菌种类无差异。随着番茄的生长，内生细菌的数量变化趋势除美星中Microbacterium spp．和Corynebacterium spp．从苗期到成熟期逐步减少外，其它种类的数量均为先增加后减少，高峰期都是在花期。不同地区种植的番茄其内生细菌的主要类群都是Bacillus spp．，Pseudomonas spp．和Agrobacterium spp．，除以上3种内生细菌外，田阳的番茄中有Xanthomonas spp．，Corynebacterium spp．，Microbacterium spp．和Erwinia spp．。与田阳的番茄内生细菌种类比较，玉林和武鸣的番茄中无Erwinia spp．，桂林的番茄中无Xanthomonas spp．，Corynebacterium spp．，Microbacterium spp．和Erwinia spp．，但发现有Enterobacter spp．。
广西大学硕士毕业论文
不同的品种植物内生细菌的种类有一定的差异，武鸣种植的番茄红宝石
一号和美星品种中，前者有xa刀th~nas sPP.，而后者无xa刀动~nas
sPP.，但是有材l’c厂obac加rl’，sPP.，玉林种植的台湾圣女和红宝石一
号番茄品种中，台湾圣女品种中有xan动，onas sPp.，和
cor夕nebac亡e厂jun7 spp.，红宝石一号品种无肠刀动，口nas sPP.和
命厂少门ebac亡e厂jum spp.，但多了汾l’c厂obacterl’，sPP.，田阳种植的番茄
中，夏辉品种中有脂n动om口刀as sPP.，和Er丽nl’a sPP.，台湾圣女品
种中无上述两个种类，但多了解croba时er/um sPp.，和
山厂夕刀ebac亡er/姗sPP.。
分离的571个菌株中，对番茄青枯病菌有拮抗作用的有14个菌株，
其中日513拮抗作用最为明显。Bs，8、日s，，、日535、日546、日m24和Bs，3对番茄
还具有促生长作用。
选用Bs，。菌株进行生物学特性测定，最适温度为31一34C，低于10’
C或高于45OC该菌停止生长，其最适pH值为7，pH值低于5或高于11
时该菌停止生长，测试的9种碳源中，Bsl。利用葡萄糖、麦芽糖、甘露醇
和山梨醇，不利用O一果糖、乳糖、O一纤维二糖、甜醇和甘油;测试的10
种氮源中，Bsl。利用OL一丙氨酸、L一天氦氨酸、蛋氨酸、甘氨酸和NaN03，
不利用L一酪氨酸、L一谷氨酸、DL一甲硫氨酸、L一甲硫氨酸、NH4GI。
根据其形态特征及生理生化反应，初步鉴定该菌株为月g厂obac亡eri，
厂adjobacte厂。
Two hundred and seventy symptomless tomato plants were collected for analysis of the population and dynamic of endophytic bacterium from Wuming, Tianyang, Yuling and guilin of Guangxi. Five hundred and seventy-one strains were obtained. The genera of tomato endophytic bacterium were Bacillus spp.,Pseudomonas spp., Xanthomonas spp., Microbacterium spp.,Corynebacterium spp. andAgrobacterium spp.. The dominated genera of tomato endophytic bacterium was Bacillus spp..
Tomato endophytic bacteria varied with the organs, growth periods ,regions and varieties. The variety of Hongbaoshi NO. 1 could not found Corynebacterium spp.in the root of tomato, and Microbacterium spp. did not live in the stems of the plants. But the kinds of endophytic bacteria had not any difference in the organs of variety of Meixing. The population of most endophytic bacteria in the growth periods of tomato increased firstly and then reduced, but Microbacterium spp. and Corynebacterium spp. reduced gradually. The main genera of endophytic bacteria were Bacillus spp.,
Pseudomonas spp. and Agrobacterium spp. in defferent regions. Excepting these three main groups, there were Xanthomonas spp. , Corynebacterium spp. , Microbacterium spp and Erwinia spp. in the tomato plants of Tianyan . By the comparision with the tomato endophytic bacteria of Tianyang, there was not Erwinia spp. in the tomato plant of Yulin and Wuming. In the tomato Plants of Guiling, Xanthomonas spp., Corynebacterium spp. , Microbacter ium spp. and Erwinia spp. were not found, but Enterobacter spp. , could live in it. There were some different in the endophytic bacteria among varieties.The variety of Hongbaoshi NO. 1 and Meixing were cultivated in Wuming. There was Xanthomonas spp. in the former variety, but later did not be found this bacteria and were found another bacterium-Microbacter ium spp. to live in it. The varieties of Taiwan Shengnu and Hongbaoshi NO.l, which were cultivated in Yulin, were also found different endophytic bacteria between them. Xanthomonas spp. and Corynebacterium spp. could be found in the variety of Taiwan Shengnu. These two bacteria could not be found in the variety of Hongbaoshi NO. 1. But Microbacter ium spp. could live in the variety of Hongbaoshi NO.l. The situation of endophytic bacteria in the variety of Xiahui and Taiwan Shengnu cultivated in Tianyang was the same as that of variety of Taiwan Shengnu and Hongbaoshi NO.l cultivated in Yulin. Xanthomonas spp. and Erwinia spp. were found in Xiahui. These two bacreria did not be found in Taiwan
shengnu, but Microbacterium spp and Corynebacterium spp. could live in the variety of Taiwan shengnu.
Fourteen of 571 isolates could inhibit the growth of Ralstonia solanacearum ,isolate Bs13 was the strongest inhibition to the pathogen among the 14 isolates. Bs18, Bsl2, Bs35, Bs46, Bm24 andBs13 had both antagonistic activity and promoting the growth of tomato.
The isolate of BsI3 was selected to test its biological characteristic. The most suitable temperature for the growth of Bs]3 was from 31 C to 34 ' C. When the temperature increased to 43 C or decreased to 10 ' C, Bs13 stoped growth. The most suitable pH for the growth of Bs13 was from 7-8. When pH increased to 11 or decreased to 5, Bs13 could not grow. Bs13 could use Maltose, Sorbitol, Mannitol, Glucose as carbon sources, it could not use D-Fructose, D-Cellobiose, Dulcitol, Glycerol and Lactose as carbon sources. L-Aspartic acid, DL-Glycine, Methionine, Glycine and NaNO3 were sutable nitrogen source for the growth of Bso, it could not use L-Tyrodsine, L-Glutamic acid, DL-Methionine, L- Methionine and NHtCl as nitrogen source. Basis on the test of morphology and biochemiology, Bso was primarily identified as Agrobacterium radiobacter .
番茄内生细菌的种群及数量随植株不同器官、生育期、地区和品种而变化。测试的红宝石品种中，根无Corynebacterium spp．，茎中无Microbacterium spp．，但在美星品种中各器官的内生细菌种类无差异。随着番茄的生长，内生细菌的数量变化趋势除美星中Microbacterium spp．和Corynebacterium spp．从苗期到成熟期逐步减少外，其它种类的数量均为先增加后减少，高峰期都是在花期。不同地区种植的番茄其内生细菌的主要类群都是Bacillus spp．，Pseudomonas spp．和Agrobacterium spp．，除以上3种内生细菌外，田阳的番茄中有Xanthomonas spp．，Corynebacterium spp．，Microbacterium spp．和Erwinia spp．。与田阳的番茄内生细菌种类比较，玉林和武鸣的番茄中无Erwinia spp．，桂林的番茄中无Xanthomonas spp．，Corynebacterium spp．，Microbacterium spp．和Erwinia spp．，但发现有Enterobacter spp．。
广西大学硕士毕业论文
不同的品种植物内生细菌的种类有一定的差异，武鸣种植的番茄红宝石
一号和美星品种中，前者有xa刀th~nas sPP.，而后者无xa刀动~nas
sPP.，但是有材l’c厂obac加rl’，sPP.，玉林种植的台湾圣女和红宝石一
号番茄品种中，台湾圣女品种中有xan动，onas sPp.，和
cor夕nebac亡e厂jun7 spp.，红宝石一号品种无肠刀动，口nas sPP.和
命厂少门ebac亡e厂jum spp.，但多了汾l’c厂obacterl’，sPP.，田阳种植的番茄
中，夏辉品种中有脂n动om口刀as sPP.，和Er丽nl’a sPP.，台湾圣女品
种中无上述两个种类，但多了解croba时er/um sPp.，和
山厂夕刀ebac亡er/姗sPP.。
分离的571个菌株中，对番茄青枯病菌有拮抗作用的有14个菌株，
其中日513拮抗作用最为明显。Bs，8、日s，，、日535、日546、日m24和Bs，3对番茄
还具有促生长作用。
选用Bs，。菌株进行生物学特性测定，最适温度为31一34C，低于10’
C或高于45OC该菌停止生长，其最适pH值为7，pH值低于5或高于11
时该菌停止生长，测试的9种碳源中，Bsl。利用葡萄糖、麦芽糖、甘露醇
和山梨醇，不利用O一果糖、乳糖、O一纤维二糖、甜醇和甘油;测试的10
种氮源中，Bsl。利用OL一丙氨酸、L一天氦氨酸、蛋氨酸、甘氨酸和NaN03，
不利用L一酪氨酸、L一谷氨酸、DL一甲硫氨酸、L一甲硫氨酸、NH4GI。
根据其形态特征及生理生化反应，初步鉴定该菌株为月g厂obac亡eri，
厂adjobacte厂。
Two hundred and seventy symptomless tomato plants were collected for analysis of the population and dynamic of endophytic bacterium from Wuming, Tianyang, Yuling and guilin of Guangxi. Five hundred and seventy-one strains were obtained. The genera of tomato endophytic bacterium were Bacillus spp.,Pseudomonas spp., Xanthomonas spp., Microbacterium spp.,Corynebacterium spp. andAgrobacterium spp.. The dominated genera of tomato endophytic bacterium was Bacillus spp..
Tomato endophytic bacteria varied with the organs, growth periods ,regions and varieties. The variety of Hongbaoshi NO. 1 could not found Corynebacterium spp.in the root of tomato, and Microbacterium spp. did not live in the stems of the plants. But the kinds of endophytic bacteria had not any difference in the organs of variety of Meixing. The population of most endophytic bacteria in the growth periods of tomato increased firstly and then reduced, but Microbacterium spp. and Corynebacterium spp. reduced gradually. The main genera of endophytic bacteria were Bacillus spp.,
Pseudomonas spp. and Agrobacterium spp. in defferent regions. Excepting these three main groups, there were Xanthomonas spp. , Corynebacterium spp. , Microbacterium spp and Erwinia spp. in the tomato plants of Tianyan . By the comparision with the tomato endophytic bacteria of Tianyang, there was not Erwinia spp. in the tomato plant of Yulin and Wuming. In the tomato Plants of Guiling, Xanthomonas spp., Corynebacterium spp. , Microbacter ium spp. and Erwinia spp. were not found, but Enterobacter spp. , could live in it. There were some different in the endophytic bacteria among varieties.The variety of Hongbaoshi NO. 1 and Meixing were cultivated in Wuming. There was Xanthomonas spp. in the former variety, but later did not be found this bacteria and were found another bacterium-Microbacter ium spp. to live in it. The varieties of Taiwan Shengnu and Hongbaoshi NO.l, which were cultivated in Yulin, were also found different endophytic bacteria between them. Xanthomonas spp. and Corynebacterium spp. could be found in the variety of Taiwan Shengnu. These two bacteria could not be found in the variety of Hongbaoshi NO. 1. But Microbacter ium spp. could live in the variety of Hongbaoshi NO.l. The situation of endophytic bacteria in the variety of Xiahui and Taiwan Shengnu cultivated in Tianyang was the same as that of variety of Taiwan Shengnu and Hongbaoshi NO.l cultivated in Yulin. Xanthomonas spp. and Erwinia spp. were found in Xiahui. These two bacreria did not be found in Taiwan
shengnu, but Microbacterium spp and Corynebacterium spp. could live in the variety of Taiwan shengnu.
Fourteen of 571 isolates could inhibit the growth of Ralstonia solanacearum ,isolate Bs13 was the strongest inhibition to the pathogen among the 14 isolates. Bs18, Bsl2, Bs35, Bs46, Bm24 andBs13 had both antagonistic activity and promoting the growth of tomato.
The isolate of BsI3 was selected to test its biological characteristic. The most suitable temperature for the growth of Bs]3 was from 31 C to 34 ' C. When the temperature increased to 43 C or decreased to 10 ' C, Bs13 stoped growth. The most suitable pH for the growth of Bs13 was from 7-8. When pH increased to 11 or decreased to 5, Bs13 could not grow. Bs13 could use Maltose, Sorbitol, Mannitol, Glucose as carbon sources, it could not use D-Fructose, D-Cellobiose, Dulcitol, Glycerol and Lactose as carbon sources. L-Aspartic acid, DL-Glycine, Methionine, Glycine and NaNO3 were sutable nitrogen source for the growth of Bso, it could not use L-Tyrodsine, L-Glutamic acid, DL-Methionine, L- Methionine and NHtCl as nitrogen source. Basis on the test of morphology and biochemiology, Bso was primarily identified as Agrobacterium radiobacter .
引文
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