草业学报 ›› 2023, Vol. 32 ›› Issue (4): 161-172.DOI: 10.11686/cyxb2022311
• 研究论文 • 上一篇
张振粉1,2(), 黄荣1,2, 姚博1,2, 张旺东3, 杨成德4, 陈秀蓉4
收稿日期:
2022-08-02
修回日期:
2022-09-05
出版日期:
2023-04-20
发布日期:
2023-01-29
通讯作者:
张振粉
作者简介:
张振粉(1984-),男,福建霞浦人,副教授,博士。 E-mail: zhangzf@gsau.edu.cn
基金资助:
Zhen-fen ZHANG1,2(), Rong HUANG1,2, Bo YAO1,2, Wang-dong ZHANG3, Cheng-de YANG4, Xiu-rong CHEN4
Received:
2022-08-02
Revised:
2022-09-05
Online:
2023-04-20
Published:
2023-01-29
Contact:
Zhen-fen ZHANG
摘要:
为了探究进口紫花苜蓿种带细菌的多样性及其对动植物的致病性,本研究从北美和欧洲共收集到紫花苜蓿种子样品34份,所有样品经室内研磨稀释分离培养,共获得39株种带细菌分离物,结合常规表型特征及16S rDNA鉴定方法确定它们的分类地位;并在室内采用菌悬液皿内发芽及盆栽接种法和腹腔注射法分别测定了21株代表细菌对供试紫花苜蓿和昆明小鼠的致病性。结果显示:1)39株细菌隶属3门15属,门分别为厚壁菌门、变形菌门和放线菌门,其中优势门为厚壁菌门;属地位的分别为芽孢杆菌属、假单胞菌属、马赛菌属、短芽孢杆菌属、欧文氏菌属、泛菌属、不动杆菌属、肠杆菌属、埃希氏肠杆菌属、假芽孢杆菌属、假节杆菌属、红球菌属、葡萄球菌属、土壤芽孢杆菌属和微杆菌属,其中优势属为芽孢杆菌属和假单胞菌属。2)红球菌属GCKH菌株仅对紫花苜蓿致病;不动杆菌属ZSR17、埃希氏肠杆菌属ZSR25和马赛菌属R1菌株仅对小鼠具有致病性;而欧文氏菌属ZF1和ZS3、泛菌属CQ10和ZS6菌株既可以引起紫花苜蓿致病,又可以引起小白鼠发病,是潜在的植物和动物跨界侵染共致病病原细菌。研究结果初步探明了欧美进口紫花苜蓿种带细菌的分类地位及其危害性,为综合防控这一类通过牧草种子携带的植物和动物独立致病细菌,以及植物和动物跨界侵染共致病病原细菌的侵入和传播奠定了理论依据。
张振粉, 黄荣, 姚博, 张旺东, 杨成德, 陈秀蓉. 欧美进口紫花苜蓿可培养种带细菌及其对动植物的致病性[J]. 草业学报, 2023, 32(4): 161-172.
Zhen-fen ZHANG, Rong HUANG, Bo YAO, Wang-dong ZHANG, Cheng-de YANG, Xiu-rong CHEN. Culturable seed-borne bacteria of lucerne imported from Europe and North America and their pathogenicity to plants and animals[J]. Acta Prataculturae Sinica, 2023, 32(4): 161-172.
编号 Code | 紫花苜蓿品种 M. sativa variety | 产地 Production region | 生产年份 Production year | 提供者 Seed provider |
---|---|---|---|---|
Lot1 | 皇冠Phabulous | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot2 | SR4030 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot3 | 巨能551 Magnum 551 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot4 | 耐盐之星Magnum Salt | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot5 | 斯贝德Spade | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot6 | 维多利亚Victory | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot7 | 巨能995 Magnum 995 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot8 | 英斯特Inster | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot9 | 威神Vision | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot10 | MT4015 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot11 | 驯鹿Caribou | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot12 | SK3010 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot13 | 巨能7 Magnum 7 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot14 | MF4020 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot15 | 巨能601 Magnum 601 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot16 | 巨能7-耐湿Magnum 7-Wet | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot17 | 巨能2 Magnum 2 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot18 | 巨能801 Magnum 801 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot19 | 旱地Handi | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot20 | 金皇后Golden empress | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot21 | 阿尔冈金Algonguin | 北美North America | 2015 | 北京克劳沃公司Beijing Clover Company |
Lot22 | 梅佐Mezzo | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot23 | 直布罗陀Gibraltar | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot24 | 超新星Super Nova | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot25 | 特蕾莎TEREZA | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot26 | 法多FADO | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot27 | 劲能4.2 Power 4.2 | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot28 | 玛格达Magda | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot29 | 富通Fortune | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot30 | 瑞诺Creno | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot31 | 萨斯卡娅Saskia | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot32 | PS2065MF | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot33 | 希模Sibemol | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot34 | 莎莎Salsa | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
表1 供试种样的来源及相关信息
Table 1 Origin and information of seed samples tested
编号 Code | 紫花苜蓿品种 M. sativa variety | 产地 Production region | 生产年份 Production year | 提供者 Seed provider |
---|---|---|---|---|
Lot1 | 皇冠Phabulous | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot2 | SR4030 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot3 | 巨能551 Magnum 551 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot4 | 耐盐之星Magnum Salt | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot5 | 斯贝德Spade | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot6 | 维多利亚Victory | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot7 | 巨能995 Magnum 995 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot8 | 英斯特Inster | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot9 | 威神Vision | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot10 | MT4015 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot11 | 驯鹿Caribou | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot12 | SK3010 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot13 | 巨能7 Magnum 7 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot14 | MF4020 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot15 | 巨能601 Magnum 601 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot16 | 巨能7-耐湿Magnum 7-Wet | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot17 | 巨能2 Magnum 2 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot18 | 巨能801 Magnum 801 | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot19 | 旱地Handi | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot20 | 金皇后Golden empress | 北美North America | 2014 | 北京克劳沃公司Beijing Clover Company |
Lot21 | 阿尔冈金Algonguin | 北美North America | 2015 | 北京克劳沃公司Beijing Clover Company |
Lot22 | 梅佐Mezzo | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot23 | 直布罗陀Gibraltar | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot24 | 超新星Super Nova | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot25 | 特蕾莎TEREZA | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot26 | 法多FADO | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot27 | 劲能4.2 Power 4.2 | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot28 | 玛格达Magda | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot29 | 富通Fortune | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot30 | 瑞诺Creno | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot31 | 萨斯卡娅Saskia | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot32 | PS2065MF | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot33 | 希模Sibemol | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
Lot34 | 莎莎Salsa | 欧洲Europe | 2015 | 丹农种子集团Dannong Seeds Bloc |
图1 紫花苜蓿种带细菌的单菌落形态特征不同字母和数字组合代表种带细菌菌株的编号。Different combinations of letters and numbers represent the number of bacterial strains.
Fig.1 Morphological characteristics of single colony of alfalfa seed-borne bacteria
编号 Code | 种带细菌总数量 Number of the colony (CFU·g-1) | 种带细菌种群数 The number of bacterial population | 优势种群 (前2位) The top two dominant bacterial population | ||
---|---|---|---|---|---|
未消毒Non-sterilized | 消毒Surface sterilized | 未消毒Non-sterilized | 消毒Surface sterilized | ||
Lot1 | 1.70×107p * | 3.70×106i | 3.33±0.33e | 2.33±0.33ab | ZSR17,ZSR18 |
Lot2 | 5.16×107j * | 3.30×106ij | 3.33±0.88e | 3.33±0.33a | ZF1,ZF23 |
Lot3 | 8.80×106q * | 1.15×106no | 3.33±0.67e | 1.33±0.33b | YCK6,YCK61 |
Lot4 | 1.43×106s | 7.00×105op | 2.33±0.33e | 2.33±0.33ab | ZSR20,ZSR21 |
Lot5 | 3.50×106r * | 1.70×105p | 3.33±0.33e | 3.33±0.88a | TM1,ZS3 |
Lot6 | 6.30×105t * | 2.20×105p | 3.33±0.88e | 3.33±0.33a | ZS6,CQ10 |
Lot7 | 1.80×107p * | 2.00×106m | 3.00±0.00e | 2.00±0.58ab | CQ10,ZS6 |
Lot8 | 2.18×108c * | 4.30×107b | 3.33±0.58e | 2.33±0.67ab | ZSR27,ZSR28 |
Lot9 | 5.15×107j * | 8.00×106g | 3.33±0.33e | 3.33±0.33a | YCK4,YCK5 |
Lot10 | 4.80×107k * | 1.60×106mn | 3.33±0.33e | 2.33±0.33ab | YCK2,ZFSH4 |
Lot11 | 6.52×107g * | 2.70×106kl | 3.67±0.33de * | 2.33±0.33ab | ZSR33,ZSR30 |
Lot12 | 3.67×107m * | 9.00×106f | 2.33±0.33e | 2.33±0.33ab | ZSR18,ZSR31 |
Lot13 | 4.26×107l * | 1.70×106mn | 3.33±0.67e | 3.33±0.33a | ZSR19,ZSR32 |
Lot14 | 6.20×107h * | 2.20×106lm | 3.33±0.67e | 3.33±0.88a | ZSR17,ZSR28 |
Lot15 | 3.78×107m * | 2.00×106m | 6.00±0.58bc * | 2.00±0.58ab | ZF1,ZF23 |
Lot16 | 3.80×107m * | 4.30×106h | 7.33±0.33ab * | 2.33±0.33ab | ZSR17,ZSR18 |
Lot17 | 5.50×107 i * | 8.00×106g | 5.33±0.33cd * | 3.33±0.33a | ZF23,QH1 |
Lot18 | 5.26×107j * | 1.60×106mn | 3.33±0.33e | 2.33±0.33ab | ZF11,CH1 |
Lot19 | 5.52×107i * | 2.70×105p | 3.67±0.67de | 2.33±0.33ab | ZF32,GCKH |
Lot20 | 1.21×108e * | 6.70×107a | 2.67±0.67e | 3.33±0.33a | GCKH,R1 |
Lot21 | 3.37×108b * | 1.16×107e | 3.00±0.58e | 2.00±0.58ab | R1,ZF1 |
Lot22 | 4.50×108a * | 1.50×107d | 2.67±0.67e | 2.33±0.33ab | ZSR18,ZSR19 |
Lot23 | 6.60×106 r * | 2.00×105p | 7.67±0.88a * | 2.00±0.58ab | YCK5,YCK51 |
Lot24 | 9.70×106q * | 3.70×105p | 3.33±0.33e | 2.33±0.88ab | ZSR29,ZSR31 |
Lot25 | 2.16×108d * | 3.30×107c | 3.33±0.33e | 3.33±0.33a | ZSR30,ZSR32 |
Lot26 | 8.78×107f * | 1.11×106no | 3.33±0.88e | 1.33±0.33b | ZFSH3,ZFSH4 |
Lot27 | 1.67×107p | 9.00×106f | 2.33±0.33e | 2.33±0.33ab | TM1,YCK4 |
Lot28 | 3.26×107n * | 1.70×106mn | 3.33±0.33e | 3.33±0.33a | TM1,QH1 |
Lot29 | 5.25×107j * | 2.20×106lm | 5.33±0.88cd * | 3.33±0.88a | ZSR22,ZSR23 |
Lot30 | 2.78×107o * | 2.00×106m | 3.00±0.58e | 2.00±0.58ab | ZS3,ZS6 |
Lot31 | 4.80×107k * | 4.30×106h | 3.33±0.33e | 2.33±0.67ab | ZS3,CQ10 |
Lot32 | 5.50×107i * | 8.00×106g | 3.33±0.88e | 3.33±0.33a | ZSR23,ZSR24 |
Lot33 | 5.26×107j * | 1.60×106mn | 3.33±0.33e | 2.33±0.33ab | ZSR25,ZSR26 |
Lot34 | 5.53×107i * | 2.70×106jk | 3.67±0.33de | 2.33±0.33ab | CQ10,ZS3 |
表2 不同产地紫花苜蓿种带细菌的种群及数量
Table 2 Seed-borne bacteria population and quantity of different varieties lucerne collected from the different locations
编号 Code | 种带细菌总数量 Number of the colony (CFU·g-1) | 种带细菌种群数 The number of bacterial population | 优势种群 (前2位) The top two dominant bacterial population | ||
---|---|---|---|---|---|
未消毒Non-sterilized | 消毒Surface sterilized | 未消毒Non-sterilized | 消毒Surface sterilized | ||
Lot1 | 1.70×107p * | 3.70×106i | 3.33±0.33e | 2.33±0.33ab | ZSR17,ZSR18 |
Lot2 | 5.16×107j * | 3.30×106ij | 3.33±0.88e | 3.33±0.33a | ZF1,ZF23 |
Lot3 | 8.80×106q * | 1.15×106no | 3.33±0.67e | 1.33±0.33b | YCK6,YCK61 |
Lot4 | 1.43×106s | 7.00×105op | 2.33±0.33e | 2.33±0.33ab | ZSR20,ZSR21 |
Lot5 | 3.50×106r * | 1.70×105p | 3.33±0.33e | 3.33±0.88a | TM1,ZS3 |
Lot6 | 6.30×105t * | 2.20×105p | 3.33±0.88e | 3.33±0.33a | ZS6,CQ10 |
Lot7 | 1.80×107p * | 2.00×106m | 3.00±0.00e | 2.00±0.58ab | CQ10,ZS6 |
Lot8 | 2.18×108c * | 4.30×107b | 3.33±0.58e | 2.33±0.67ab | ZSR27,ZSR28 |
Lot9 | 5.15×107j * | 8.00×106g | 3.33±0.33e | 3.33±0.33a | YCK4,YCK5 |
Lot10 | 4.80×107k * | 1.60×106mn | 3.33±0.33e | 2.33±0.33ab | YCK2,ZFSH4 |
Lot11 | 6.52×107g * | 2.70×106kl | 3.67±0.33de * | 2.33±0.33ab | ZSR33,ZSR30 |
Lot12 | 3.67×107m * | 9.00×106f | 2.33±0.33e | 2.33±0.33ab | ZSR18,ZSR31 |
Lot13 | 4.26×107l * | 1.70×106mn | 3.33±0.67e | 3.33±0.33a | ZSR19,ZSR32 |
Lot14 | 6.20×107h * | 2.20×106lm | 3.33±0.67e | 3.33±0.88a | ZSR17,ZSR28 |
Lot15 | 3.78×107m * | 2.00×106m | 6.00±0.58bc * | 2.00±0.58ab | ZF1,ZF23 |
Lot16 | 3.80×107m * | 4.30×106h | 7.33±0.33ab * | 2.33±0.33ab | ZSR17,ZSR18 |
Lot17 | 5.50×107 i * | 8.00×106g | 5.33±0.33cd * | 3.33±0.33a | ZF23,QH1 |
Lot18 | 5.26×107j * | 1.60×106mn | 3.33±0.33e | 2.33±0.33ab | ZF11,CH1 |
Lot19 | 5.52×107i * | 2.70×105p | 3.67±0.67de | 2.33±0.33ab | ZF32,GCKH |
Lot20 | 1.21×108e * | 6.70×107a | 2.67±0.67e | 3.33±0.33a | GCKH,R1 |
Lot21 | 3.37×108b * | 1.16×107e | 3.00±0.58e | 2.00±0.58ab | R1,ZF1 |
Lot22 | 4.50×108a * | 1.50×107d | 2.67±0.67e | 2.33±0.33ab | ZSR18,ZSR19 |
Lot23 | 6.60×106 r * | 2.00×105p | 7.67±0.88a * | 2.00±0.58ab | YCK5,YCK51 |
Lot24 | 9.70×106q * | 3.70×105p | 3.33±0.33e | 2.33±0.88ab | ZSR29,ZSR31 |
Lot25 | 2.16×108d * | 3.30×107c | 3.33±0.33e | 3.33±0.33a | ZSR30,ZSR32 |
Lot26 | 8.78×107f * | 1.11×106no | 3.33±0.88e | 1.33±0.33b | ZFSH3,ZFSH4 |
Lot27 | 1.67×107p | 9.00×106f | 2.33±0.33e | 2.33±0.33ab | TM1,YCK4 |
Lot28 | 3.26×107n * | 1.70×106mn | 3.33±0.33e | 3.33±0.33a | TM1,QH1 |
Lot29 | 5.25×107j * | 2.20×106lm | 5.33±0.88cd * | 3.33±0.88a | ZSR22,ZSR23 |
Lot30 | 2.78×107o * | 2.00×106m | 3.00±0.58e | 2.00±0.58ab | ZS3,ZS6 |
Lot31 | 4.80×107k * | 4.30×106h | 3.33±0.33e | 2.33±0.67ab | ZS3,CQ10 |
Lot32 | 5.50×107i * | 8.00×106g | 3.33±0.88e | 3.33±0.33a | ZSR23,ZSR24 |
Lot33 | 5.26×107j * | 1.60×106mn | 3.33±0.33e | 2.33±0.33ab | ZSR25,ZSR26 |
Lot34 | 5.53×107i * | 2.70×106jk | 3.67±0.33de | 2.33±0.33ab | CQ10,ZS3 |
菌株 Strain | 大小 Size (μm) | 革兰氏染色 Gram stain | NCBI中相似菌株 Similar strain | 相似度 Similarity (%) | 登录号 Genebank accession |
---|---|---|---|---|---|
ZSR17 | 0.37~0.60×0.78~2.28 | - | 不动杆菌属Acinetobacter sp. | 99 | KY312517 |
ZSR18 | 0.60~1.10×0.89~2.34 | - | 假单胞菌属Pseudomonas sp. | 99 | KY312518 |
ZSR19 | 0.19~0.57×0.35~3.61 | - | 肠杆菌属Enterobacter sp. | 99 | KY436203 |
ZSR20 | 0.31~0.66×0.78~3.05 | + | 萎缩芽孢杆菌Bacillus atrophaeus | 99 | KY436204 |
ZSR21 | 0.34~0.69×0.43~1.65 | + | 微杆菌属Microbacterium sp. | 99 | KY436205 |
ZSR22 | 0.38~0.84×0.54~2.77 | + | 奇异假芽孢杆菌Fictibacillus barbaricus | 99 | KY436206 |
ZSR23 | 0.43~0.89×0.82~4.19 | + | 耐寒短芽孢杆菌Brevibacillus frigoritolerans | 99 | KY436207 |
ZSR24 | 0.38~0.69×0.78~2.34 | - | 浅黄假单胞菌Pseudomonas luteola | 99 | KY436208 |
ZSR25 | 0.26~0.49×0.42~1.12 | - | 埃希氏菌属Escherichia sp. | 99 | KY436209 |
ZSR26 | 0.60~1.20×1.65~6.67 | + | 内生芽孢杆菌Bacillus endophyticus | 99 | KY436210 |
ZSR27 | 0.69~1.39×1.57~11.54 | + | 嗜盐土地芽孢杆菌Terribacillus saccharophilus | 99 | KY436211 |
ZSR28 | 0.25~0.56×0.69~2.86 | + | 简单芽孢杆菌Bacillus simplex | 100 | KY436212 |
ZSR29 | 0.54~1.20×2.04~5.96 | + | 简单芽孢杆菌B. simplex | 99 | KY436213 |
ZSR30 | 0.53~1.03×1.84×7.73 | + | 简单芽孢杆菌B. simplex | 99 | KY436214 |
ZSR31 | 0.62~1.33×2.72~7.57 | + | 奇异假芽孢杆菌F. barbaricus | 99 | KY436215 |
ZSR32 | 0.46~0.94×1.52~5.77 | + | 简单芽孢杆菌B. simplex | 99 | KY436216 |
ZSR33 | 0.60~1.62×1.46~3.46 | + | 耐寒短芽孢杆菌B. frigoritolerans | 99 | KY436217 |
ZF9 | 0.73~1.17×3.84~11.09 | + | 蜡样芽孢杆菌Bacillus cereus | 99 | KX784910 |
ZFSH2 | 0.38~0.73×1.30~5.74 | + | 短小芽孢杆菌Bacillus pumilus | 99 | KX784911 |
ZF1 | 0.57~1.40×1.62~7.00 | - | 欧文氏菌属Erwinia sp. | 99 | KX784912 |
ZF23 | 1.28~3.83×2.10~4.78 | + | 地衣芽孢杆菌Bacillus licheniformis | 99 | KX784914 |
QH1 | 0.65~1.53×1.84~7.32 | + | 蜡样芽孢杆菌B. cereus | 99 | KX784915 |
ZF11 | 0.50~1.03×0.57~1.82 | + | 蜡样芽孢杆菌B. cereus | 99 | KX784916 |
CH1 | 0.49~0.73×1.49~3.50 | + | 葡萄球菌属Staphylococcus sp. | 99 | KX784917 |
ZF32 | 0.45~0.85×1.27~4.23 | + | 枯草芽孢杆菌Bacillus subtilis | 99 | KX784918 |
GCKH | 0.34~0.99×0.49~1.50 | + | 红球菌属Rhodococcus sp. | 99 | KX784919 |
R1 | 0.36~0.85×0.68~2.75 | - | 变异马赛菌Massilia varians | 99 | KX784920 |
YCK61 | 0.36~0.69×0.61~2.14 | + | 蜡样芽孢杆菌B. cereus | 99 | KX784921 |
YCK6 | 0.30~0.58×0.57~1.40 | + | 氧化假节杆菌Pseudarthrobacter oxydans | 99 | KX784922 |
YCK51 | 0.37~0.66×0.99~2.69 | + | 硬结假节杆菌Pseudarthrobacter scleromae | 99 | KX784923 |
YCK5 | 0.36~0.85×0.69~1.87 | - | 短杆状马赛菌Massilia brevitalea | 99 | KX784924 |
YCK4 | 0.46~1.21×1.02~3.20 | - | 栖稻假单胞菌Pseudomonas oryzihabitans | 99 | KX784925 |
YCK2 | 0.31~0.55×0.43~1.63 | + | 土壤短芽孢杆菌Brevibacillus agri | 99 | KX784926 |
ZFSH4 | 0.34~0.55×0.62~2.95 | - | 恶臭假单胞菌Pseudomonas putida | 99 | KX784927 |
ZFSH3 | 0.26~0.60×0.94~3.54 | - | 蒂莫内马赛菌Massilia timonae | 99 | KX676495 |
TM1 | 0.46~0.86×0.84~1.96 | - | 铜绿假单胞菌Pseudomonas aeruginosa | 99 | KY305014 |
ZS3 | 0.31~0.55×0.43~1.63 | - | 大黄欧文氏菌Erwinia rhapontici | 99 | KY495782 |
ZS6 | 0.34~0.55×0.62~2.65 | - | 成团泛菌Pantoea agglomerans | 99 | KT454095 |
CQ10 | 0.46~0.86×0.84~1.96 | - | 泛菌属Pantoea sp. | 99 | KT355269 |
表3 39株紫花苜蓿种带细菌菌株的鉴定
Table 3 Identification of 39 bacterial strains from alfalfa seeds
菌株 Strain | 大小 Size (μm) | 革兰氏染色 Gram stain | NCBI中相似菌株 Similar strain | 相似度 Similarity (%) | 登录号 Genebank accession |
---|---|---|---|---|---|
ZSR17 | 0.37~0.60×0.78~2.28 | - | 不动杆菌属Acinetobacter sp. | 99 | KY312517 |
ZSR18 | 0.60~1.10×0.89~2.34 | - | 假单胞菌属Pseudomonas sp. | 99 | KY312518 |
ZSR19 | 0.19~0.57×0.35~3.61 | - | 肠杆菌属Enterobacter sp. | 99 | KY436203 |
ZSR20 | 0.31~0.66×0.78~3.05 | + | 萎缩芽孢杆菌Bacillus atrophaeus | 99 | KY436204 |
ZSR21 | 0.34~0.69×0.43~1.65 | + | 微杆菌属Microbacterium sp. | 99 | KY436205 |
ZSR22 | 0.38~0.84×0.54~2.77 | + | 奇异假芽孢杆菌Fictibacillus barbaricus | 99 | KY436206 |
ZSR23 | 0.43~0.89×0.82~4.19 | + | 耐寒短芽孢杆菌Brevibacillus frigoritolerans | 99 | KY436207 |
ZSR24 | 0.38~0.69×0.78~2.34 | - | 浅黄假单胞菌Pseudomonas luteola | 99 | KY436208 |
ZSR25 | 0.26~0.49×0.42~1.12 | - | 埃希氏菌属Escherichia sp. | 99 | KY436209 |
ZSR26 | 0.60~1.20×1.65~6.67 | + | 内生芽孢杆菌Bacillus endophyticus | 99 | KY436210 |
ZSR27 | 0.69~1.39×1.57~11.54 | + | 嗜盐土地芽孢杆菌Terribacillus saccharophilus | 99 | KY436211 |
ZSR28 | 0.25~0.56×0.69~2.86 | + | 简单芽孢杆菌Bacillus simplex | 100 | KY436212 |
ZSR29 | 0.54~1.20×2.04~5.96 | + | 简单芽孢杆菌B. simplex | 99 | KY436213 |
ZSR30 | 0.53~1.03×1.84×7.73 | + | 简单芽孢杆菌B. simplex | 99 | KY436214 |
ZSR31 | 0.62~1.33×2.72~7.57 | + | 奇异假芽孢杆菌F. barbaricus | 99 | KY436215 |
ZSR32 | 0.46~0.94×1.52~5.77 | + | 简单芽孢杆菌B. simplex | 99 | KY436216 |
ZSR33 | 0.60~1.62×1.46~3.46 | + | 耐寒短芽孢杆菌B. frigoritolerans | 99 | KY436217 |
ZF9 | 0.73~1.17×3.84~11.09 | + | 蜡样芽孢杆菌Bacillus cereus | 99 | KX784910 |
ZFSH2 | 0.38~0.73×1.30~5.74 | + | 短小芽孢杆菌Bacillus pumilus | 99 | KX784911 |
ZF1 | 0.57~1.40×1.62~7.00 | - | 欧文氏菌属Erwinia sp. | 99 | KX784912 |
ZF23 | 1.28~3.83×2.10~4.78 | + | 地衣芽孢杆菌Bacillus licheniformis | 99 | KX784914 |
QH1 | 0.65~1.53×1.84~7.32 | + | 蜡样芽孢杆菌B. cereus | 99 | KX784915 |
ZF11 | 0.50~1.03×0.57~1.82 | + | 蜡样芽孢杆菌B. cereus | 99 | KX784916 |
CH1 | 0.49~0.73×1.49~3.50 | + | 葡萄球菌属Staphylococcus sp. | 99 | KX784917 |
ZF32 | 0.45~0.85×1.27~4.23 | + | 枯草芽孢杆菌Bacillus subtilis | 99 | KX784918 |
GCKH | 0.34~0.99×0.49~1.50 | + | 红球菌属Rhodococcus sp. | 99 | KX784919 |
R1 | 0.36~0.85×0.68~2.75 | - | 变异马赛菌Massilia varians | 99 | KX784920 |
YCK61 | 0.36~0.69×0.61~2.14 | + | 蜡样芽孢杆菌B. cereus | 99 | KX784921 |
YCK6 | 0.30~0.58×0.57~1.40 | + | 氧化假节杆菌Pseudarthrobacter oxydans | 99 | KX784922 |
YCK51 | 0.37~0.66×0.99~2.69 | + | 硬结假节杆菌Pseudarthrobacter scleromae | 99 | KX784923 |
YCK5 | 0.36~0.85×0.69~1.87 | - | 短杆状马赛菌Massilia brevitalea | 99 | KX784924 |
YCK4 | 0.46~1.21×1.02~3.20 | - | 栖稻假单胞菌Pseudomonas oryzihabitans | 99 | KX784925 |
YCK2 | 0.31~0.55×0.43~1.63 | + | 土壤短芽孢杆菌Brevibacillus agri | 99 | KX784926 |
ZFSH4 | 0.34~0.55×0.62~2.95 | - | 恶臭假单胞菌Pseudomonas putida | 99 | KX784927 |
ZFSH3 | 0.26~0.60×0.94~3.54 | - | 蒂莫内马赛菌Massilia timonae | 99 | KX676495 |
TM1 | 0.46~0.86×0.84~1.96 | - | 铜绿假单胞菌Pseudomonas aeruginosa | 99 | KY305014 |
ZS3 | 0.31~0.55×0.43~1.63 | - | 大黄欧文氏菌Erwinia rhapontici | 99 | KY495782 |
ZS6 | 0.34~0.55×0.62~2.65 | - | 成团泛菌Pantoea agglomerans | 99 | KT454095 |
CQ10 | 0.46~0.86×0.84~1.96 | - | 泛菌属Pantoea sp. | 99 | KT355269 |
图2 39株紫花苜蓿种带细菌基于16S rDNA序列的系统发育树分支距离表示各序列之间的成对进化距离。Branch distance represents the pairwise evolutionary distance between sequences.
Fig.2 Phylogenetic tree of 39 seed-borne bacteria of alfalfa based on 16S rDNA sequences
菌株 Strain | 紫花苜蓿Lucerne | 小白鼠 Mice | |
---|---|---|---|
皿内发芽Petri plate germination | 盆栽Pot culture | ||
ZSR17 | - | - | +厌食、寒颤Anorexia, shivering |
ZSR18 | - | - | - |
ZSR19 | - | - | - |
ZSR20 | - | - | - |
ZSR21 | - | - | - |
ZSR22 | - | - | - |
ZSR23 | - | - | - |
ZSR25 | - | - | +腹泻、寒颤Diarrhea, shivering |
ZSR27 | - | - | - |
ZF9 | - | - | - |
ZF1 | +萎蔫Wilt | +萎蔫Wilt | +厌食、寒颤Anorexia, shivering |
CH1 | - | - | - |
GCKH | +叶斑Leaf spot | +叶斑Leaf spot | - |
R1 | - | - | +腹泻、寒颤Diarrhea, shivering |
YCK6 | - | - | - |
YCK5 | - | - | +厌食、寒颤Anorexia, shivering |
ZFSH3 | - | - | +厌食、寒颤Anorexia, shivering |
TM1 | - | - | +厌食、寒颤Anorexia, shivering |
ZS3 | +萎蔫Wilt | +萎蔫Wilt | +厌食、寒颤Anorexia, shivering |
ZS6 | +叶斑Leaf spot | +叶斑Leaf spot | +厌食、寒颤Anorexia, shivering |
CQ10 | +叶斑Leaf spot | +叶斑Leaf spot | +厌食、寒颤Anorexia, shivering |
表4 供试代表细菌菌株的致病性
Table 4 The pathogenicity of tested bacterial strains
菌株 Strain | 紫花苜蓿Lucerne | 小白鼠 Mice | |
---|---|---|---|
皿内发芽Petri plate germination | 盆栽Pot culture | ||
ZSR17 | - | - | +厌食、寒颤Anorexia, shivering |
ZSR18 | - | - | - |
ZSR19 | - | - | - |
ZSR20 | - | - | - |
ZSR21 | - | - | - |
ZSR22 | - | - | - |
ZSR23 | - | - | - |
ZSR25 | - | - | +腹泻、寒颤Diarrhea, shivering |
ZSR27 | - | - | - |
ZF9 | - | - | - |
ZF1 | +萎蔫Wilt | +萎蔫Wilt | +厌食、寒颤Anorexia, shivering |
CH1 | - | - | - |
GCKH | +叶斑Leaf spot | +叶斑Leaf spot | - |
R1 | - | - | +腹泻、寒颤Diarrhea, shivering |
YCK6 | - | - | - |
YCK5 | - | - | +厌食、寒颤Anorexia, shivering |
ZFSH3 | - | - | +厌食、寒颤Anorexia, shivering |
TM1 | - | - | +厌食、寒颤Anorexia, shivering |
ZS3 | +萎蔫Wilt | +萎蔫Wilt | +厌食、寒颤Anorexia, shivering |
ZS6 | +叶斑Leaf spot | +叶斑Leaf spot | +厌食、寒颤Anorexia, shivering |
CQ10 | +叶斑Leaf spot | +叶斑Leaf spot | +厌食、寒颤Anorexia, shivering |
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