盐碱胁迫下一株促进苜蓿生长的细菌筛选与鉴定
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摘要
为改良和利用盐碱地,本研究以大庆草原盐碱土为材料,在Na_2CO_3和NaHCO_3终浓度分别为50、100 mmol·L~(-1),pH 8.5的条件下对其中耐盐碱细菌进行分离,并在CAS铁载体检测培养基、ADF培养基、PKO无机磷培养基等功能性培养基上对菌株的促生功能进行检测;利用16S rDNA序列分析法进行菌株的菌种鉴定,在pH 8.5,25 mmol·L~(-1) Na_2CO_3胁迫下测试菌株对苜蓿种子发芽率、发芽势及幼苗生长的影响。结果表明:筛选得到的菌株S11具有产植物激素IAA和产ACC脱氨酶功能;通过对菌株16S rDNA序列进行分析,初步鉴定菌株S11为堀越氏芽孢杆菌(Bacillus horikoshii)。盐碱胁迫条件下,浸种接菌处理苜蓿种子后,种子发芽率和发芽势分别提高了15%和10%,幼苗根长、株高、鲜重以及叶绿素含量分别增加了34.78%、12.5%、27.3%和43.30%,这证明菌株S11具有缓解盐碱胁迫促进苜蓿生长的功能。
In order to ameliorate and utilize saline-alkali soil, the saline-alkali soil was used to separate the saline-alkali-tolerant bacteria under pH 8.5, 50 mmol·L~(-1) Na_2CO_3 and 100 mmol·L~(-1) NaHCO_3. The growth-promoting function of the strain was tested by CAS, ADF, PKO and other functional medium. The 16 S rDNA sequence analysis was used to identify the strains. The effect of the bacteria on the alfalfa seed germination rate, germination potential and seedling growth were investigated under pH 8.5 and 25 mmol·L~(-1) Na_2CO_3. The results showed that strain S11 could produce IAA and ACC deaminase, and it was identified as Bacillus horikoshii. Under the saline-alkali condition and after inoculation of strain S11, alfalfa seed germination rate and germination potential were increased by 15% and 10%, respectively. The root length, plant height, fresh biomass, and chlorophyll content were increased by 34.78%、12.5%、27.3% and 43.30%, respectively. This proved that strain S11 could relieve saline-alkali stress and promote the growth of alfalfa.
In order to ameliorate and utilize saline-alkali soil, the saline-alkali soil was used to separate the saline-alkali-tolerant bacteria under pH 8.5, 50 mmol·L~(-1) Na_2CO_3 and 100 mmol·L~(-1) NaHCO_3. The growth-promoting function of the strain was tested by CAS, ADF, PKO and other functional medium. The 16 S rDNA sequence analysis was used to identify the strains. The effect of the bacteria on the alfalfa seed germination rate, germination potential and seedling growth were investigated under pH 8.5 and 25 mmol·L~(-1) Na_2CO_3. The results showed that strain S11 could produce IAA and ACC deaminase, and it was identified as Bacillus horikoshii. Under the saline-alkali condition and after inoculation of strain S11, alfalfa seed germination rate and germination potential were increased by 15% and 10%, respectively. The root length, plant height, fresh biomass, and chlorophyll content were increased by 34.78%、12.5%、27.3% and 43.30%, respectively. This proved that strain S11 could relieve saline-alkali stress and promote the growth of alfalfa.
引文
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[14] 荣良燕,姚拓,赵桂琴,等.产铁载体PGPR菌筛选及其对病原菌的拮抗作用[J].植物保护,2011,(1):59-64.
[15] 郑鹏.ACC脱氨酶根际促生菌的分离鉴定及其对干旱胁迫下玉米甜菜碱代谢的影响[D].咸阳:西北农林科技大学,2015.
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[18] 李波,于海龙.苏打盐碱胁迫下不同来源苜蓿品种抗盐性综合评价[J].草地学报,2017,25(1):204-208.
[19] 张飞鹏,林志伟,张振南,等.大庆盐碱土中微生物的分离及芽孢杆菌的鉴定[J].黑龙江八一农垦大学学报,2011,23(5):6-8.
[20] 颜宏,赵伟,尹尚军,等.羊草对不同盐碱胁迫的生理响应[J].草业学报,2006,15(6):49-55.
[21] 刘冠一,刘艳玲,刘博文,等.含ACC脱氨酶活性的复合菌株提高苜蓿抗盐碱能力研究[J].核农学报,2017,(5):1022-1028.
[22] Ullah A,Heng S,Munis M F H,et al.Phytoremediation of heavy metals assisted by plant growth promoting (PGP) bacteria:a review[J].Environmental and Experimental Botany,2015,117:28-40.
[23] 殷丽华,徐鹏,柯希望,等.混合盐碱胁迫对绿豆种子萌发与幼苗生长发育的影响[J].黑龙江八一农垦大学学报,2015,(5):40-43.
[24] Ramadoss D,Lakkineni V K,Bose P,et al.Mitigation of salt stress in wheat seedlings by halotolerant bacteria isolated from saline habitats[J].Springer plus,2013,2(1):1-7.
[25] Bharti N,Barnawal D,Awasthi A,et al.Plant growth promoting rhizobacteria alleviate salinity induced negative effects on growth,oil content and physiological status in Mentha arvensis[J].Acta Physiologiae Plantarum,2014,36(1):45-60.
[26] Xie Y,Han S,Li X,et al.Ameliorates of salt stress on bermuda grass by the fungus aspergillus aculeatus[J].Molecular plant-microbe interactions,2017,30(3):245-254.
[2] 武祎,田雨,张红香,等.盐、碱胁迫与温度对黄花苜蓿种子发芽的影响[J].草业科学,2015(11):1847-1853.
[3] Parray J A,Jan S,Kamili A N,et al.Current perspectives on plant growth-promoting rhizobacteria[J].Journal of Plant Growth Regulation,2016,35(3):877-902.
[4] 蔺吉祥,高战武,王颖,等.盐碱胁迫对紫花苜蓿种子发芽的协同影响[J].草地学报,2014,22(2):312-318.
[5] 赵琦,包玉英.混合盐碱胁迫下丛枝菌根真菌对紫花苜蓿生长及2种酚酸含量的影响[J].西北植物学报,2015,35(9):1829-1836.
[6] 张福海,罗会,崔松山,等.耐盐促生菌剂对盐碱环境下枸杞生长的影响[J].园艺与种苗,2017,(7):33-35.
[7] 庞学兵,王朝阳,王爱英,等.根际促生菌对棉苗盐碱胁迫的缓解效应[J].西北农业学报,2017,(1):101-109.
[8] Poli A,Esposito E,Orlando P,et al.Halomonas alkaliantarctica sp nov.,isolated from saline lake Cape Russell in Antarctica,an alkalophilic moderately halophilic,exopolysaccharide-producing bacterium[J].Systematic and Applied Microbiology,2007,30(1):31-38.
[9] 石伟.极端盐碱土壤细菌的分离筛选及抗盐特性研究[D].哈尔滨:东北林业大学,2011.
[10] Gupta R,Singal R,Shankar A,et al.A modified plate assay for screening phosphate solubilizing microorganisms[J].Journal of General & Applied Microbiology,2006,40(3):255-260.
[11] 李艳星,郭平毅,孙建光.块根块茎类作物内生固氮菌分离鉴定、系统发育与促生特性[J].中国农业科学,2017,(1):104-122.
[12] Glickmann E,Dessaux Y.A critical examination of the specificity of the salkowski reagent for indolic compounds produced by phytopathogenic bacteria[J].Applied & Environmental Microbiology,1995,61(2):793-796.
[13] 刘莎莎.植物根际促生菌互作提高苜蓿抗盐碱能力研究[D].哈尔滨:哈尔滨师范大学,2016.
[14] 荣良燕,姚拓,赵桂琴,等.产铁载体PGPR菌筛选及其对病原菌的拮抗作用[J].植物保护,2011,(1):59-64.
[15] 郑鹏.ACC脱氨酶根际促生菌的分离鉴定及其对干旱胁迫下玉米甜菜碱代谢的影响[D].咸阳:西北农林科技大学,2015.
[16] 詹寿发,卢丹妮,毛花英,等.2株溶磷、解钾与产IAA的内生真菌菌株的筛选、鉴定及促生作用研究[J].中国土壤与肥料,2017,(3):142-151.
[17] 尉晶.浅析盐碱胁迫对燕麦种子萌发和幼苗的影响[J].黑龙江八一农垦大学学报,2017,(5):13-15,88.
[18] 李波,于海龙.苏打盐碱胁迫下不同来源苜蓿品种抗盐性综合评价[J].草地学报,2017,25(1):204-208.
[19] 张飞鹏,林志伟,张振南,等.大庆盐碱土中微生物的分离及芽孢杆菌的鉴定[J].黑龙江八一农垦大学学报,2011,23(5):6-8.
[20] 颜宏,赵伟,尹尚军,等.羊草对不同盐碱胁迫的生理响应[J].草业学报,2006,15(6):49-55.
[21] 刘冠一,刘艳玲,刘博文,等.含ACC脱氨酶活性的复合菌株提高苜蓿抗盐碱能力研究[J].核农学报,2017,(5):1022-1028.
[22] Ullah A,Heng S,Munis M F H,et al.Phytoremediation of heavy metals assisted by plant growth promoting (PGP) bacteria:a review[J].Environmental and Experimental Botany,2015,117:28-40.
[23] 殷丽华,徐鹏,柯希望,等.混合盐碱胁迫对绿豆种子萌发与幼苗生长发育的影响[J].黑龙江八一农垦大学学报,2015,(5):40-43.
[24] Ramadoss D,Lakkineni V K,Bose P,et al.Mitigation of salt stress in wheat seedlings by halotolerant bacteria isolated from saline habitats[J].Springer plus,2013,2(1):1-7.
[25] Bharti N,Barnawal D,Awasthi A,et al.Plant growth promoting rhizobacteria alleviate salinity induced negative effects on growth,oil content and physiological status in Mentha arvensis[J].Acta Physiologiae Plantarum,2014,36(1):45-60.
[26] Xie Y,Han S,Li X,et al.Ameliorates of salt stress on bermuda grass by the fungus aspergillus aculeatus[J].Molecular plant-microbe interactions,2017,30(3):245-254.