苜蓿根瘤菌与苜蓿品种共生匹配优良组合筛选的研究
|
摘要
苜蓿是一种具有较高经济价值和营养价值的多年生豆科牧草。种植苜蓿既可以为畜禽提供优质的绿色蛋白饲料,又可以改良土壤,提高肥力,对生态环境具有深远的意义。提高苜蓿产量有效的方法是给苜蓿接种与之相匹配的根瘤菌。本研究是以中国农业微生物菌种保藏管理中心国家农业菌库中7株推广应用的苜蓿根瘤菌与6个苜蓿品种进行的共生匹配的初步探讨,并对7株根瘤菌采用生理生化特性反应、Biolog细菌鉴定系统和16S rDNA序列分析方法进行系统鉴定。通过根瘤菌与不同苜蓿品种在无氮水培营养液中进行结瘤试验和有效性分析,初筛出与对照相比共生匹配效果较好根瘤菌6株,苜蓿品种5个,共15个组合。将这15个组合进行土壤盆栽试验,分析了5个苜蓿品种接种6株根瘤菌的共生固氮生长效应,筛选出每一个苜蓿品种最佳共生效应的根瘤菌菌株,初步确立苜蓿根瘤菌与苜蓿品种共生匹配的优良组合5个,将复筛后的优良组合进行田间效果试验,为苜蓿根瘤菌更好地推广应用奠定基础。
试验结果如下:
1.将7株根瘤菌与6个苜蓿品种通过滤纸桥法,在无氮水培营养液中进行苜蓿根瘤菌与苜蓿品种的共生匹配结瘤试验,比较各个匹配组合与对照的差异,初筛出与对照相比较结瘤效果较好的共生匹配组合15个。
2.土壤盆栽试验模拟田间试验的土壤环境,采用土壤不灭菌进行根瘤菌与苜蓿品种的共生匹配试验,对初筛出的较好根瘤菌与苜蓿组合在土壤中的适应性、结瘤能力和固氮有效性进行研究。结果表明,不同苜蓿品种接种根瘤菌ACCC17674与对照相比在结瘤率、根瘤数量、植株高度、植株鲜重、植株干重和全氮上有显著提高,且与其它菌株间差异显著,说明ACCC17674具有较强广谱性。共筛出共生匹配优良组合5个,分别是阿尔冈金苜蓿与ACCC17674、金皇后苜蓿与ACCC17674、牧歌苜蓿与ACCC17674、淮阴苜蓿与ACCC17674和保定苜蓿与ACCC17674。
3.将复筛出的5个优良组合进行田间小区试验,结果表明,不同苜蓿品种接种ACCC17674与对照相比结瘤率、植株高度、干重、分枝数、全氮和产草量显著提高。最终得到3个共生匹配的优良组合,进一步验证了ACCC17674是一株广谱菌株,可以进一步推广应用到实际生产。
4.在七株供试根瘤菌的生理生化特性试验中,在耐酸碱性方面,菌株ACCC17674和ACCC17675耐酸碱的pH值生长范围在4.0-9.5之间,耐盐性可达4.5%左右,说明两株菌在田间应用时,能够适应土壤pH值的较强变化和耐受高盐土壤环境。
5.利用Biolog细菌鉴定系统和16S rDNA序列分析对供试7株根瘤菌进行系统鉴定时,7株菌均属于苜蓿中华根瘤菌(Sinorhizobium melilot),其鉴定结果与传统鉴定方法的鉴定相一致。在传统生理生化鉴定方法的基础上,利用当代先进的分析鉴定系统对根瘤菌的鉴定更系统更全面。
The alfalfa is one kind of perennial leguminous forage with high economic value and nutritional value. The alfalfa can not only provide the high quality green protein feed for the domesticated fowl, but also improve soil and enhance the fertility, which is profoundly significant to the ecological environment. The effective method of increasing the alfalfa yield is the vaccination of the matching nodule bacteria on it. In the present study, the symbiotic of 7 strains of alfalfa rhizobia from Agricultural Culture Collection of China (ACCC) and 6 alfalfa varieties were investigated preliminarily and 7 Sinorhizobium meliloti were identified systemically by the reaction characteristic of the physiological biochemistry, the Biolog system of bacterim identification and the 16S rDNA sequential analysis method.
Through nodulation experiments and effectiveness analysis of nodule bacterium and different alfalfa varieties in the non-nitrogen liquid medium, compared with the control, 6 rhizobia and 5 alfalfa varieties with better symbiotic matching effects were screened and 15 combinations were developed. The pot trial was conducted with these 15 combinations and the symbiotic nitrogen fixation growth effects of 5 alfalfa varieties vaccinated with 6 rhizobia were studied, and the rhizobia with the best symbiotic effect on ach alfalfa variety was selected and 5 combinations of the alfalfa nodule bacteria and the alfalfa variety with good symbiotic matching were screened, followed by field trial of the 5 combinations, which will be the foundation. of promoting the application of alfalfa nodule bacteria.
The results were as following:
1. The symbiotic matching experiment between 7 rhizobia and 6 alfalfa varieties were conducted by filter paper bridge culture method, in the non-nitrogen liquid medium culture and the differences between each matching combination and the control were compared. 15 combinations with better effect of symbiotic matching were screened.
2. By the simulation of the field soil environment in pot trial, the symbiotic matching test between rhizobia and the alfalfa varieties was conducted with antiseptic soil, and the adaptability of screened rhizobia and alfalfa combination, nodulation ability and the efficiency of nitrogen fixation were studied. The results indicated that nodulation rate, nodule quantity, the plant height, the plant fresh weight and dry weight, the total nitrogen when rhizobium ACCC17674 vaccinated on the different alfalfa varieties were significantly increased, compared with control and significant differenced between ACCC17674 and the other strains were found. It is explained that ACCC17674 is a broad-spectrum strain. Finally, 5 combinations were screening respectively Algonquin, Golden Empress, Madrigal, HuaiYin and Baoding with ACCC17674 .
3. A field plot trial was conducted with 5 screened good combinations. The results showed that nodulation rate, the plant height, branch number, the total nitrogen and yield were significantly increased and 3 combinations with good symbiotic matching were obtained, which had the potential of application in practical production.It is explained that ACCC17674 is a broad-spectrum strain.
4. In the physiological biochemistry characteristics reaction test of 7 rhizobia, strains ACCC17674 and ACCC17675 could grow with the pH value of 4.0-9.5 and 4.5% salt, and it suggested that in field application, the 2 strains could be adapted to the soil of with high pH value change and high salt content, with the good adaptability to environment and the higher tolerance.
5. 7 Sinorhizobium meliloti identified by Biolog identification system and the 16S rDNA sequence analysis, the results were consistent with the traditional methods. Therefore, the advcanced analysis methods were more systematic and comprehensive on the basis of traditional methods in bacterial identification.
试验结果如下:
1.将7株根瘤菌与6个苜蓿品种通过滤纸桥法,在无氮水培营养液中进行苜蓿根瘤菌与苜蓿品种的共生匹配结瘤试验,比较各个匹配组合与对照的差异,初筛出与对照相比较结瘤效果较好的共生匹配组合15个。
2.土壤盆栽试验模拟田间试验的土壤环境,采用土壤不灭菌进行根瘤菌与苜蓿品种的共生匹配试验,对初筛出的较好根瘤菌与苜蓿组合在土壤中的适应性、结瘤能力和固氮有效性进行研究。结果表明,不同苜蓿品种接种根瘤菌ACCC17674与对照相比在结瘤率、根瘤数量、植株高度、植株鲜重、植株干重和全氮上有显著提高,且与其它菌株间差异显著,说明ACCC17674具有较强广谱性。共筛出共生匹配优良组合5个,分别是阿尔冈金苜蓿与ACCC17674、金皇后苜蓿与ACCC17674、牧歌苜蓿与ACCC17674、淮阴苜蓿与ACCC17674和保定苜蓿与ACCC17674。
3.将复筛出的5个优良组合进行田间小区试验,结果表明,不同苜蓿品种接种ACCC17674与对照相比结瘤率、植株高度、干重、分枝数、全氮和产草量显著提高。最终得到3个共生匹配的优良组合,进一步验证了ACCC17674是一株广谱菌株,可以进一步推广应用到实际生产。
4.在七株供试根瘤菌的生理生化特性试验中,在耐酸碱性方面,菌株ACCC17674和ACCC17675耐酸碱的pH值生长范围在4.0-9.5之间,耐盐性可达4.5%左右,说明两株菌在田间应用时,能够适应土壤pH值的较强变化和耐受高盐土壤环境。
5.利用Biolog细菌鉴定系统和16S rDNA序列分析对供试7株根瘤菌进行系统鉴定时,7株菌均属于苜蓿中华根瘤菌(Sinorhizobium melilot),其鉴定结果与传统鉴定方法的鉴定相一致。在传统生理生化鉴定方法的基础上,利用当代先进的分析鉴定系统对根瘤菌的鉴定更系统更全面。
The alfalfa is one kind of perennial leguminous forage with high economic value and nutritional value. The alfalfa can not only provide the high quality green protein feed for the domesticated fowl, but also improve soil and enhance the fertility, which is profoundly significant to the ecological environment. The effective method of increasing the alfalfa yield is the vaccination of the matching nodule bacteria on it. In the present study, the symbiotic of 7 strains of alfalfa rhizobia from Agricultural Culture Collection of China (ACCC) and 6 alfalfa varieties were investigated preliminarily and 7 Sinorhizobium meliloti were identified systemically by the reaction characteristic of the physiological biochemistry, the Biolog system of bacterim identification and the 16S rDNA sequential analysis method.
Through nodulation experiments and effectiveness analysis of nodule bacterium and different alfalfa varieties in the non-nitrogen liquid medium, compared with the control, 6 rhizobia and 5 alfalfa varieties with better symbiotic matching effects were screened and 15 combinations were developed. The pot trial was conducted with these 15 combinations and the symbiotic nitrogen fixation growth effects of 5 alfalfa varieties vaccinated with 6 rhizobia were studied, and the rhizobia with the best symbiotic effect on ach alfalfa variety was selected and 5 combinations of the alfalfa nodule bacteria and the alfalfa variety with good symbiotic matching were screened, followed by field trial of the 5 combinations, which will be the foundation. of promoting the application of alfalfa nodule bacteria.
The results were as following:
1. The symbiotic matching experiment between 7 rhizobia and 6 alfalfa varieties were conducted by filter paper bridge culture method, in the non-nitrogen liquid medium culture and the differences between each matching combination and the control were compared. 15 combinations with better effect of symbiotic matching were screened.
2. By the simulation of the field soil environment in pot trial, the symbiotic matching test between rhizobia and the alfalfa varieties was conducted with antiseptic soil, and the adaptability of screened rhizobia and alfalfa combination, nodulation ability and the efficiency of nitrogen fixation were studied. The results indicated that nodulation rate, nodule quantity, the plant height, the plant fresh weight and dry weight, the total nitrogen when rhizobium ACCC17674 vaccinated on the different alfalfa varieties were significantly increased, compared with control and significant differenced between ACCC17674 and the other strains were found. It is explained that ACCC17674 is a broad-spectrum strain. Finally, 5 combinations were screening respectively Algonquin, Golden Empress, Madrigal, HuaiYin and Baoding with ACCC17674 .
3. A field plot trial was conducted with 5 screened good combinations. The results showed that nodulation rate, the plant height, branch number, the total nitrogen and yield were significantly increased and 3 combinations with good symbiotic matching were obtained, which had the potential of application in practical production.It is explained that ACCC17674 is a broad-spectrum strain.
4. In the physiological biochemistry characteristics reaction test of 7 rhizobia, strains ACCC17674 and ACCC17675 could grow with the pH value of 4.0-9.5 and 4.5% salt, and it suggested that in field application, the 2 strains could be adapted to the soil of with high pH value change and high salt content, with the good adaptability to environment and the higher tolerance.
5. 7 Sinorhizobium meliloti identified by Biolog identification system and the 16S rDNA sequence analysis, the results were consistent with the traditional methods. Therefore, the advcanced analysis methods were more systematic and comprehensive on the basis of traditional methods in bacterial identification.
引文
1.陈廷伟,非豆科作物固氮研究进展。中国农业科技出版社,1994年第二版,3-9
2.陈华癸,樊庆生.微生物学.农业出版社. 1985.150
3.陈文新.中国共生固氮研究五十年[M].南京农业出版社,1987,134～136.
4.陈文新,汪恩涛,陈文峰.根瘤菌一豆科植物共生多样性与地理环境的关系[J].中国农业科学. 2004.37 (1): 81～86
5.陈雯莉,李阜棣,周俊初.用叶绿素评价快生型大豆根瘤菌的共生有效性.华中农业大学学报,1996,(1 S):46～50.
6.崔国文.紫花苜蓿的国内外生产现状及市场分析预测.黑龙江畜牧兽医2001, (8):19.
7.东秀珠,蔡妙英常见细菌系统鉴定手册.科学出版社。2001
8.樊庆笙,接种根瘤菌的结瘤率取决于菌系的专适性和寄主的遗传性。中国微生物学会论文摘要汇编,1979.187
9.葛诚主编,微生物肥料生产应用基础。中国农业科技出版社,2000年9月第一版,29-58
10.高振生,马其东,刘志强等.沿海滩涂地区苜蓿根瘤菌接种方法和效果的研究[[J].草地学报,1996. ( 4): 288～293
11.耿华珠编.中国苜蓿[M].中国农业出版社北京: 1995.10
12.诸葛健,王正祥.工业微生物实验技术手册.中国轻工业出版社。1994.117～120
13.虎彪.紫花苜蓿和红三叶草接种根瘤菌试验研究.西南民族学院学报,1997.23(2):159～161.
14.胡济生,豆科植物固氮效率大田调查方法。中国农业科学,1964.8:20～23
15.洪绂曾.积极稳步发展中国的苜蓿产业。动物科学与动物医学,2001, 18 (4 ):1～3.
16.联合国粮食及农业组织.生物固氮技术手册.罗马,1983.
17.李世雄,王彦荣,孙建华.中国苜蓿品种种子产量性状的遗传多样性[J] .草业学报2003, 12( 1) : 23 ~29.
18.李红梅.植物的共生固氮体系[[J].生物学杂志,2004.21 (5 ): 60-61
19.李凤兰,王宏,董卫民等.紫花苜蓿根瘤菌接种菌种筛选试验研究[J].草与畜杂志,1998. (1):17～18
20.刘宏生.辽宁省豆科结瘤植物及其根瘤菌资源调查[J].生态学杂志,2000 . 6: 62～64.
21.刘惠琴.根瘤菌肥料及其应用[[J].土壤肥料. 1994. (1): 37~40
22.苗永清.优良牧草一紫花苜蓿.山西水土保持科技,1998(2):30～32.
23.孟庆杰,王光学.2004.生物固氮及其应用.生物学教学.29 (5): 6～7
24.梅笑漫.生物固氮研究中的几个热点问题.[J].生物学杂志. 2002.19 (4): 7～9
25.马玉珍,史清亮,王静.山西省根瘤菌资源的种类与共生效应[J].山西农业大学学报,1998,(1): 65-69.
26.马晓彤,宁国赞,刘慧琴,我国苜蓿根瘤菌与苜蓿共生固氮优良组合研究进展及前景。中国苜蓿发展大会.第二届。北京2003:74～76
27. M.奥巴托等著,蒋有绎,周枫译.共生固氮技术手册.北京:科学出版社,1987,47~53
28.宁国赞,刘慧琴,马晓彤.豆科根瘤菌及其应用技术.中原农民出版社.1998.21
29.宁国赞,刘惠琴,马晓彤.生物固氮技术在退耕还林还草中的应用.中国草地,2001.4, 70
30.宁国赞,刘惠琴,马晓彤.中国豆科牧草根瘤菌资源的采集保藏及利用,草地学报,1999,7(2):165～172
31.宁国赞,刘惠琴,马晓彤.中国豆科牧草根瘤菌资源的采集保藏及利用[[J].草地学报,1999 (2):165～172.
32.宁国赞,李元芳,刘慧琴等紫花苜蓿接种根瘤菌的效果。草业科学,1992,9(1):50~51
33.沈世华,荆玉祥.2003.中国生物固氮研究现状和进展[J].科学通报.48 (6) 35-40
34.孙鸿良.西北地区农牧业发展方向探讨[J].草业学报,2003, 12( 4) : 1 ~6施莱杰,H.Cx.普通微生物学.复旦大学出版社. 1990.386
35.土壤农业化学常规分析方法.科学出版社.1983.
36.王培.中国草地科学进展(C).第二届年会暨学术讨论会文集.北京:中国农业大学出版社.1998.4: 132～135
37.王素英,李润花,刘新成等.西藏部分地区豆科植物根瘤菌资源的初步调查[J].西北农林科技大学学报(自然科学版),2002 . 1: 33～37.
38.王素英,李新锁,陈文新等.河北豆科植物根瘤菌资源的初步调查研究[[J].天津师大学报(自然科学版),2000 . (3):32～36.
39.王卫卫,胡正海,关桂兰等.日一肃、宁夏部分地区根瘤菌资源及其共生固氮特性[[J].自然资源学报,2002 . (1): 48～54.
40.王静,马玉珍,史清亮等.山西根瘤菌资源多样性与特异性研究[J].应用与环境生物学报,1999,1: 79～84.
41.王索柱,叶莉,董卫民等.豆科牧草的根瘤菌接种试验.中国牧业通迅,2002,(7):59～60.
42.韦革宏,陈文新,朱铭莪.陕甘宁地区根瘤菌的16SrDNA PCR-RFLP分析[J].农业生物技术学报,2000,4: 333～336.
43.韦革宏,陈文新,朱铭莪.陕甘宁地区根瘤菌数值分类与DNA同源性分析[J].应用与环境生物学报,1999,1: 73～78.
44.向述荣,任莉.豆科牧草根瘤菌的研究进展与应用概况[J].草与畜杂志,1996, (1 ):5～7
45.云锦凤,孙启忠。抓住机遇开创我国苜蓿产业化新局面。第二届中国苜蓿发展大会。2003,北京。4～6
46.俞大绂,李季伦.微生物学.科学出版社. 1985.
47.姚拓.饲用燕麦和小麦根际根瘤菌特性研究及其生物菌肥的初步研制[D]: 2002【.博士学位论文1.兰州:甘肃农业大学
48.喻文虎,杨鹏翼,贾德荣等.红豆草、紫花苜蓿根瘤菌接种研究.草业科学,1995,12(3):22～25.
49. [英]J L,斯普朗特著,刘永定译,黎尚豪校.Ill氮生物学期工作总结.北京:农业出版社,1985,10:183～194.
50.朱奇,陈彦,生物固氮在我国农业生产中的研究现状及发展对策。微生物学杂志,2003,23(5):40～44
51.张宪武,大豆根瘤菌在不同土壤上接种效果试验。土壤微生物学集刊,第一号:52～57
52.周平贞等.豆科植物结瘤试验—水培法介绍.中国油料,第二期,I 979,50~62.
53.中国科学院微生物研究所细菌分类组编著,一般细菌常用鉴定方法。科学出版社,1978
54.中国微生物学菌种保藏管理委员会编.中国菌种目录.北京:机械工业出版社,1995.
55.曾昭海,隋新华,胡跃高等.紫花苜蓿-根瘤菌高效共生体筛选及田间作用效果.草业学报. 2004 (10): 95-100
56.曾昭海,隋新华,胡跃高等.紫花苜蓿根瘤菌高效共生体筛选及其田间作用效果研究IR].北京:第二届中国苜蓿发展大会2003.68~70
57. Allen E.K. and Allen O.N. The scope of nodulation in the leguminosae.In Recen Advances in Botany, Vol.l.Proceedings of the Ninth International Botanical Congress. 1961.PP.585～588
58. Ayneabeba Adamu, Fassil Assefa, Mariam A.H, et al. Studies of rhizobium inoculation and fertilizer treatment on growth and production of faba bean (Vicia faba) in some‘yield-depleted' and `yield sustained'regions of Semien Shewa[J]. Sinet Ethiopian J Sci, 2001, 24 (2): 197--211
59. Barber, L. D., Joern, B. C., Volenec, J. J, et al. Supplemental nitrogen effects on alfalfa regrowth and nitrogen mobilization from roots. Crop Science .1996,36:1217-1223
60. Bruce A Hungate, Peter D Stiling, Paul Dijkstra, et al. COz elicits long-term decline in nitrogen fixation[J].Science, 2004, 304 (5675): 1291
61. Catherine dhont, Yves castonguay, Paul nadea. Nitrogen reserves, spring regrowth and wintersurvival of field-grown alfalfa (Medicago sativa L) defoliated in the autumn. Annals of Botany,2006,97:109-120
62. Chen W, Wang E, Wang S, et al. Characteristics of Rhizobium tianshanese sp. nov. a moderately and slowly growing root nodule bacterium isolated from an arid saline environment in Xinjiang, people's Republic of China[J]. Int. J. Syst. Bacteriol., 1995, 45(1): 153-159
63. Craig Edminster,Don miller,Jim Moutry.美国的苜蓿产业。动物科学与动物医学,2001,18(4):58～61
64. Dhont C, Castonguay Y, Nadeau P, Be’langer C. Chalifour F P. Alfalfa root nitrogen reserves and regrowth potential in response to fall harvests .Crop Science, 2003,43: 181-194
65. Diane M. Hebb, Alan E. Richardson, R. Reid et al. PCR as an ecological tool to determine the establishment and persistence of Rhizobium strains introduced into the field as seed inoculant. Aust.J. Agric. Res. . 1998, 49: 923-934
66. Duhigg P,Melton B,Baltensperger A.Selection for acetylene reducetion rates in“Mesilla”alfalfa[J].Crop Science,1978,18:813~816
67. Gibson, A. H.. Genetic variation in the effictiveness of modulation of Lucerne varieties. Aust. J.Agric. Res.. 1962, 13: 388～399
68. Gasser, H. P, Guy, M. O., Sikora, L. Efficiency of Rhizobium meliloti strains and their effects on alfalfa cultivars. Can. J. Plant Sci. 1972, 52: 444～448
69. Gresshoff P.M. Nitrogen fixation:achievements and objectives[M]. New York:1986.
70. Hardataon,G,Heichel, G. H., Vance, C. P et目.1 Evaluation of alfalfa and lthizobium meliloti for compatibility in modulation and nodule effectiveness. 1981, Crop Science. 21: 562-566
71. Heichel G H. Barnes D K. et al. Evaluation of alfalfa and Rhizohium meliloti for compatibilily in nodulalion and nodule effectiveness [J].Crop.Sci.,1981.21:562～567
72. Heidstra R. Genetic and pHylogeny of rhizobia isolated from tropical tree legumes (PHD thesis) University of Helsinki, 1997
73. Heuer HM, Baker KP, Smalla K,Wellington EM Analysis of actinomycete communities by specific amplification of genes encoding 16S rDNA and gel-eletrophoretic separation in denaturing gradients. Appl Environ Microbiol. 1997 63: 3233～3241
74. Janice E.T.et a1. Influense of the size of indigenous Rhizobial populations on establishment and symbiotic performance of introduced Rhizobia on field-grown Legumes. Appl.Environ.Microbiol. 1991.57(1):1928
75. LongSR.Rhizobium-legume nodulation:life together in the underground.Cell.1989,56:203~214
76. Matsuda A, Moreira FM de S, Siqueira JO, et al. Tolerance of rhizobia genera from different origins to zinc,copper and cadmium[J]. Pesquida Agropecuaria Brasileira, 2002, 37 (3): 343-355
77. Michael D, Hoy Kenneth. J .Mor. Alfalfa Yield and Quality as Influenced by establishment method. Agronomy Journal ,2002 ,94(1-2):1163-1170
78. Mirza N A,Bohlool B B,Somasergaran P. Non-destructive chlorophyll assay for screening of strains of Bradychixobium japonicum. Soil Biol Biochemj,1990,22(2):203～207.
79. Newton W.E. et a1.1981.Nitrogen fixation: its scope and importance in nitrogen fixation ed:Hr11er A. and W.E. Newton (Plenum Press) P.1.
80. Nutman, P S. Varietal differences in the nodulation of subterranean clover. Aust. J. A.Res.1967, 18: 381
81. Paul E F and Ford B W. Nitrogen fertilizers: Meeting Contemporary Challenges[J].AMBO. 2002.31 (2): 169～176
82. Perret X, Steahelin C, Broughton W J. Molecular basis of symbiotic promiscuity. Microbiology and Molecular Biology Reviews. 2000.64:180～201
83. Provorov N. A., Sainmnarov U. B., Tanriverdiev TA and Simarov B.V The contributions of plant and bacteria genotypes in the growth and nitrogen accumulation of the inoculated alfalfa. Plant and Soi1. 1994.164.213～219
84. Rao Suba N.S. Soil Microbiology.Fourth edition of soil microorganisma and plant growth..Science Publishers, Inc. USA .1999.
85. Richardson A E., Viccars E A., Watson J M. and Gibson A H. Differentiation of Rhizobium strains using the ploymerase chain reaction with random and directed primers. Soil Biology&Biochemistry ,1995,27:515-524.
86. Sawada H, H leki, H Oyaizu and S Matsumoto. Proposal for rejection ofAgrobacterium tumefaciens and revised descriptions for the genus Agrobacterium and for Agrobacteium rhizogenes. Int J Syst Bacteriol,1993, 43:694-702.
87. Segundo E,Martinez-Aarca F,Dillewijn P,et a1. Characterisation of symbiotically efficient alfalfa-nodulating rhizobia isolated from acid soils of Argentina and Uruguay[J].FEMS Microbiology Ecology. 1999.28(2):169- 176
88. Socolow,R.H. Nitrogen management and the future of food:Lessons from the management of energy and carbon.Proc.Natl.Achd.Sci.USA.1999,96:6001～6008
89. Sprent,J.I.,Sprent,P. Nitrogen fixation Organisms.Pure and Applied Aspects.Sprent J.I.and Sprent P (eds).Chapman and Hall,London,United Kingdom.1990
90. Sharma, P. B., Ajit Singh, Rangil Singh. Symbiotic nitrogen fixation by winter legumes. Acomparative efficiency between an Indian and an Australian variety of legume, Medicago saliva.Indian J. Agric. Res. 1973, 7(3-4):159~163
91. Tan G Y,T an W K. Interaction between alfalfa cultivars and Rhiaobiurn strains for nitronen fixation. Theoretical and Applied Genetics[J].1986,71:724~29.
92. Tan G Y,Tan W K,Net assim ilation rate and relative nitrogen assimilateion rate in relation to the dry matter production of alfalfa cultivars[J].Plant Soil,1981,59:185~192
93. Temprano, M. Albareda et a1. Survival of Several Rhizobium/Bradyrhizobium Strains on Different Inoculation and InoculatedSeeds,lnt Microbia1. 2002.5:81～86
94. Zhang T, Fan X,Hanada S, Kamagata Y, Fang H Bacillus macauensis sp. nov., a long-chain bacterium isolated from a drinking water supply. Int J Syst Evol Microbiol. 2006, 56: 349～353
95. Zabran H H.Rhizobium-Legume symbiosis and nitrogen fixation under severe conditions and in arid climate[J].Microbiol.Mol.Biol.Rev,1999,(63):968～989.
96. Zakhia F., De Lajudie, P. Taxonomy of rhizobium.Agronomie. 2001.21:569-576
2.陈华癸,樊庆生.微生物学.农业出版社. 1985.150
3.陈文新.中国共生固氮研究五十年[M].南京农业出版社,1987,134～136.
4.陈文新,汪恩涛,陈文峰.根瘤菌一豆科植物共生多样性与地理环境的关系[J].中国农业科学. 2004.37 (1): 81～86
5.陈雯莉,李阜棣,周俊初.用叶绿素评价快生型大豆根瘤菌的共生有效性.华中农业大学学报,1996,(1 S):46～50.
6.崔国文.紫花苜蓿的国内外生产现状及市场分析预测.黑龙江畜牧兽医2001, (8):19.
7.东秀珠,蔡妙英常见细菌系统鉴定手册.科学出版社。2001
8.樊庆笙,接种根瘤菌的结瘤率取决于菌系的专适性和寄主的遗传性。中国微生物学会论文摘要汇编,1979.187
9.葛诚主编,微生物肥料生产应用基础。中国农业科技出版社,2000年9月第一版,29-58
10.高振生,马其东,刘志强等.沿海滩涂地区苜蓿根瘤菌接种方法和效果的研究[[J].草地学报,1996. ( 4): 288～293
11.耿华珠编.中国苜蓿[M].中国农业出版社北京: 1995.10
12.诸葛健,王正祥.工业微生物实验技术手册.中国轻工业出版社。1994.117～120
13.虎彪.紫花苜蓿和红三叶草接种根瘤菌试验研究.西南民族学院学报,1997.23(2):159～161.
14.胡济生,豆科植物固氮效率大田调查方法。中国农业科学,1964.8:20～23
15.洪绂曾.积极稳步发展中国的苜蓿产业。动物科学与动物医学,2001, 18 (4 ):1～3.
16.联合国粮食及农业组织.生物固氮技术手册.罗马,1983.
17.李世雄,王彦荣,孙建华.中国苜蓿品种种子产量性状的遗传多样性[J] .草业学报2003, 12( 1) : 23 ~29.
18.李红梅.植物的共生固氮体系[[J].生物学杂志,2004.21 (5 ): 60-61
19.李凤兰,王宏,董卫民等.紫花苜蓿根瘤菌接种菌种筛选试验研究[J].草与畜杂志,1998. (1):17～18
20.刘宏生.辽宁省豆科结瘤植物及其根瘤菌资源调查[J].生态学杂志,2000 . 6: 62～64.
21.刘惠琴.根瘤菌肥料及其应用[[J].土壤肥料. 1994. (1): 37~40
22.苗永清.优良牧草一紫花苜蓿.山西水土保持科技,1998(2):30～32.
23.孟庆杰,王光学.2004.生物固氮及其应用.生物学教学.29 (5): 6～7
24.梅笑漫.生物固氮研究中的几个热点问题.[J].生物学杂志. 2002.19 (4): 7～9
25.马玉珍,史清亮,王静.山西省根瘤菌资源的种类与共生效应[J].山西农业大学学报,1998,(1): 65-69.
26.马晓彤,宁国赞,刘慧琴,我国苜蓿根瘤菌与苜蓿共生固氮优良组合研究进展及前景。中国苜蓿发展大会.第二届。北京2003:74～76
27. M.奥巴托等著,蒋有绎,周枫译.共生固氮技术手册.北京:科学出版社,1987,47~53
28.宁国赞,刘慧琴,马晓彤.豆科根瘤菌及其应用技术.中原农民出版社.1998.21
29.宁国赞,刘惠琴,马晓彤.生物固氮技术在退耕还林还草中的应用.中国草地,2001.4, 70
30.宁国赞,刘惠琴,马晓彤.中国豆科牧草根瘤菌资源的采集保藏及利用,草地学报,1999,7(2):165～172
31.宁国赞,刘惠琴,马晓彤.中国豆科牧草根瘤菌资源的采集保藏及利用[[J].草地学报,1999 (2):165～172.
32.宁国赞,李元芳,刘慧琴等紫花苜蓿接种根瘤菌的效果。草业科学,1992,9(1):50~51
33.沈世华,荆玉祥.2003.中国生物固氮研究现状和进展[J].科学通报.48 (6) 35-40
34.孙鸿良.西北地区农牧业发展方向探讨[J].草业学报,2003, 12( 4) : 1 ~6施莱杰,H.Cx.普通微生物学.复旦大学出版社. 1990.386
35.土壤农业化学常规分析方法.科学出版社.1983.
36.王培.中国草地科学进展(C).第二届年会暨学术讨论会文集.北京:中国农业大学出版社.1998.4: 132～135
37.王素英,李润花,刘新成等.西藏部分地区豆科植物根瘤菌资源的初步调查[J].西北农林科技大学学报(自然科学版),2002 . 1: 33～37.
38.王素英,李新锁,陈文新等.河北豆科植物根瘤菌资源的初步调查研究[[J].天津师大学报(自然科学版),2000 . (3):32～36.
39.王卫卫,胡正海,关桂兰等.日一肃、宁夏部分地区根瘤菌资源及其共生固氮特性[[J].自然资源学报,2002 . (1): 48～54.
40.王静,马玉珍,史清亮等.山西根瘤菌资源多样性与特异性研究[J].应用与环境生物学报,1999,1: 79～84.
41.王索柱,叶莉,董卫民等.豆科牧草的根瘤菌接种试验.中国牧业通迅,2002,(7):59～60.
42.韦革宏,陈文新,朱铭莪.陕甘宁地区根瘤菌的16SrDNA PCR-RFLP分析[J].农业生物技术学报,2000,4: 333～336.
43.韦革宏,陈文新,朱铭莪.陕甘宁地区根瘤菌数值分类与DNA同源性分析[J].应用与环境生物学报,1999,1: 73～78.
44.向述荣,任莉.豆科牧草根瘤菌的研究进展与应用概况[J].草与畜杂志,1996, (1 ):5～7
45.云锦凤,孙启忠。抓住机遇开创我国苜蓿产业化新局面。第二届中国苜蓿发展大会。2003,北京。4～6
46.俞大绂,李季伦.微生物学.科学出版社. 1985.
47.姚拓.饲用燕麦和小麦根际根瘤菌特性研究及其生物菌肥的初步研制[D]: 2002【.博士学位论文1.兰州:甘肃农业大学
48.喻文虎,杨鹏翼,贾德荣等.红豆草、紫花苜蓿根瘤菌接种研究.草业科学,1995,12(3):22～25.
49. [英]J L,斯普朗特著,刘永定译,黎尚豪校.Ill氮生物学期工作总结.北京:农业出版社,1985,10:183～194.
50.朱奇,陈彦,生物固氮在我国农业生产中的研究现状及发展对策。微生物学杂志,2003,23(5):40～44
51.张宪武,大豆根瘤菌在不同土壤上接种效果试验。土壤微生物学集刊,第一号:52～57
52.周平贞等.豆科植物结瘤试验—水培法介绍.中国油料,第二期,I 979,50~62.
53.中国科学院微生物研究所细菌分类组编著,一般细菌常用鉴定方法。科学出版社,1978
54.中国微生物学菌种保藏管理委员会编.中国菌种目录.北京:机械工业出版社,1995.
55.曾昭海,隋新华,胡跃高等.紫花苜蓿-根瘤菌高效共生体筛选及田间作用效果.草业学报. 2004 (10): 95-100
56.曾昭海,隋新华,胡跃高等.紫花苜蓿根瘤菌高效共生体筛选及其田间作用效果研究IR].北京:第二届中国苜蓿发展大会2003.68~70
57. Allen E.K. and Allen O.N. The scope of nodulation in the leguminosae.In Recen Advances in Botany, Vol.l.Proceedings of the Ninth International Botanical Congress. 1961.PP.585～588
58. Ayneabeba Adamu, Fassil Assefa, Mariam A.H, et al. Studies of rhizobium inoculation and fertilizer treatment on growth and production of faba bean (Vicia faba) in some‘yield-depleted' and `yield sustained'regions of Semien Shewa[J]. Sinet Ethiopian J Sci, 2001, 24 (2): 197--211
59. Barber, L. D., Joern, B. C., Volenec, J. J, et al. Supplemental nitrogen effects on alfalfa regrowth and nitrogen mobilization from roots. Crop Science .1996,36:1217-1223
60. Bruce A Hungate, Peter D Stiling, Paul Dijkstra, et al. COz elicits long-term decline in nitrogen fixation[J].Science, 2004, 304 (5675): 1291
61. Catherine dhont, Yves castonguay, Paul nadea. Nitrogen reserves, spring regrowth and wintersurvival of field-grown alfalfa (Medicago sativa L) defoliated in the autumn. Annals of Botany,2006,97:109-120
62. Chen W, Wang E, Wang S, et al. Characteristics of Rhizobium tianshanese sp. nov. a moderately and slowly growing root nodule bacterium isolated from an arid saline environment in Xinjiang, people's Republic of China[J]. Int. J. Syst. Bacteriol., 1995, 45(1): 153-159
63. Craig Edminster,Don miller,Jim Moutry.美国的苜蓿产业。动物科学与动物医学,2001,18(4):58～61
64. Dhont C, Castonguay Y, Nadeau P, Be’langer C. Chalifour F P. Alfalfa root nitrogen reserves and regrowth potential in response to fall harvests .Crop Science, 2003,43: 181-194
65. Diane M. Hebb, Alan E. Richardson, R. Reid et al. PCR as an ecological tool to determine the establishment and persistence of Rhizobium strains introduced into the field as seed inoculant. Aust.J. Agric. Res. . 1998, 49: 923-934
66. Duhigg P,Melton B,Baltensperger A.Selection for acetylene reducetion rates in“Mesilla”alfalfa[J].Crop Science,1978,18:813~816
67. Gibson, A. H.. Genetic variation in the effictiveness of modulation of Lucerne varieties. Aust. J.Agric. Res.. 1962, 13: 388～399
68. Gasser, H. P, Guy, M. O., Sikora, L. Efficiency of Rhizobium meliloti strains and their effects on alfalfa cultivars. Can. J. Plant Sci. 1972, 52: 444～448
69. Gresshoff P.M. Nitrogen fixation:achievements and objectives[M]. New York:1986.
70. Hardataon,G,Heichel, G. H., Vance, C. P et目.1 Evaluation of alfalfa and lthizobium meliloti for compatibility in modulation and nodule effectiveness. 1981, Crop Science. 21: 562-566
71. Heichel G H. Barnes D K. et al. Evaluation of alfalfa and Rhizohium meliloti for compatibilily in nodulalion and nodule effectiveness [J].Crop.Sci.,1981.21:562～567
72. Heidstra R. Genetic and pHylogeny of rhizobia isolated from tropical tree legumes (PHD thesis) University of Helsinki, 1997
73. Heuer HM, Baker KP, Smalla K,Wellington EM Analysis of actinomycete communities by specific amplification of genes encoding 16S rDNA and gel-eletrophoretic separation in denaturing gradients. Appl Environ Microbiol. 1997 63: 3233～3241
74. Janice E.T.et a1. Influense of the size of indigenous Rhizobial populations on establishment and symbiotic performance of introduced Rhizobia on field-grown Legumes. Appl.Environ.Microbiol. 1991.57(1):1928
75. LongSR.Rhizobium-legume nodulation:life together in the underground.Cell.1989,56:203~214
76. Matsuda A, Moreira FM de S, Siqueira JO, et al. Tolerance of rhizobia genera from different origins to zinc,copper and cadmium[J]. Pesquida Agropecuaria Brasileira, 2002, 37 (3): 343-355
77. Michael D, Hoy Kenneth. J .Mor. Alfalfa Yield and Quality as Influenced by establishment method. Agronomy Journal ,2002 ,94(1-2):1163-1170
78. Mirza N A,Bohlool B B,Somasergaran P. Non-destructive chlorophyll assay for screening of strains of Bradychixobium japonicum. Soil Biol Biochemj,1990,22(2):203～207.
79. Newton W.E. et a1.1981.Nitrogen fixation: its scope and importance in nitrogen fixation ed:Hr11er A. and W.E. Newton (Plenum Press) P.1.
80. Nutman, P S. Varietal differences in the nodulation of subterranean clover. Aust. J. A.Res.1967, 18: 381
81. Paul E F and Ford B W. Nitrogen fertilizers: Meeting Contemporary Challenges[J].AMBO. 2002.31 (2): 169～176
82. Perret X, Steahelin C, Broughton W J. Molecular basis of symbiotic promiscuity. Microbiology and Molecular Biology Reviews. 2000.64:180～201
83. Provorov N. A., Sainmnarov U. B., Tanriverdiev TA and Simarov B.V The contributions of plant and bacteria genotypes in the growth and nitrogen accumulation of the inoculated alfalfa. Plant and Soi1. 1994.164.213～219
84. Rao Suba N.S. Soil Microbiology.Fourth edition of soil microorganisma and plant growth..Science Publishers, Inc. USA .1999.
85. Richardson A E., Viccars E A., Watson J M. and Gibson A H. Differentiation of Rhizobium strains using the ploymerase chain reaction with random and directed primers. Soil Biology&Biochemistry ,1995,27:515-524.
86. Sawada H, H leki, H Oyaizu and S Matsumoto. Proposal for rejection ofAgrobacterium tumefaciens and revised descriptions for the genus Agrobacterium and for Agrobacteium rhizogenes. Int J Syst Bacteriol,1993, 43:694-702.
87. Segundo E,Martinez-Aarca F,Dillewijn P,et a1. Characterisation of symbiotically efficient alfalfa-nodulating rhizobia isolated from acid soils of Argentina and Uruguay[J].FEMS Microbiology Ecology. 1999.28(2):169- 176
88. Socolow,R.H. Nitrogen management and the future of food:Lessons from the management of energy and carbon.Proc.Natl.Achd.Sci.USA.1999,96:6001～6008
89. Sprent,J.I.,Sprent,P. Nitrogen fixation Organisms.Pure and Applied Aspects.Sprent J.I.and Sprent P (eds).Chapman and Hall,London,United Kingdom.1990
90. Sharma, P. B., Ajit Singh, Rangil Singh. Symbiotic nitrogen fixation by winter legumes. Acomparative efficiency between an Indian and an Australian variety of legume, Medicago saliva.Indian J. Agric. Res. 1973, 7(3-4):159~163
91. Tan G Y,T an W K. Interaction between alfalfa cultivars and Rhiaobiurn strains for nitronen fixation. Theoretical and Applied Genetics[J].1986,71:724~29.
92. Tan G Y,Tan W K,Net assim ilation rate and relative nitrogen assimilateion rate in relation to the dry matter production of alfalfa cultivars[J].Plant Soil,1981,59:185~192
93. Temprano, M. Albareda et a1. Survival of Several Rhizobium/Bradyrhizobium Strains on Different Inoculation and InoculatedSeeds,lnt Microbia1. 2002.5:81～86
94. Zhang T, Fan X,Hanada S, Kamagata Y, Fang H Bacillus macauensis sp. nov., a long-chain bacterium isolated from a drinking water supply. Int J Syst Evol Microbiol. 2006, 56: 349～353
95. Zabran H H.Rhizobium-Legume symbiosis and nitrogen fixation under severe conditions and in arid climate[J].Microbiol.Mol.Biol.Rev,1999,(63):968～989.
96. Zakhia F., De Lajudie, P. Taxonomy of rhizobium.Agronomie. 2001.21:569-576
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |