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构树(Broussonetia papyrifera (Linnaeus) L’Héritier ex Ventenat)是一种具有重要经济价值的多年生乔木,属于桑科(Moraceae)构属(Broussonetia)。根据Angiosperm Phylogeny Group III分类系统,又属固氮分支[1]。近年来,新品种杂交构树华构1号因抗逆性强、生长速度快、产量高、粗蛋白质含量高的优点,被用作优质的木本饲料来源在全国20多个省(市)推广种植;但在甘肃等西北干旱与半干旱地区,杂交构树的生长存在产量低、翌年返青率低等问题[2]。针对以上问题,研究人员一方面开展了选育适宜甘肃冷凉地区的高产抗寒性品系的工作;另一方面,鉴于研究表明杂交构树高含量的粗蛋白来源和速生可能与其内生和根际共生的共生固氮或联合固氮菌有密切关系[3],可通过从杂交构树根际土壤中筛选本土固氮菌微生物再回接的方式,提高菌肥效果,达到增产的目的。因此,根据土壤环境情况分离筛选杂交构树优良固氮菌株,是研制菌肥最基础的前期工作。本研究旨在从甘肃地区种植的杂交构树根际分离筛选获得固氮菌株,并对其固氮、促生、溶磷、生物防治等功能进行研究,可为进一步开发甘肃地区杂交构树微生物氮肥提供菌种资源。
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经16S rDNA测序比对发现,从3个试验区杂交构树根际共分离到10株固氮菌,有5株菌为3地共有,分属同1菌株(表1)。与该属模式菌株及最高相似性菌株的16S rDNA基因序列构建的系统发育树(图1)表明:10株菌分属8个属9个种,其中,3地共有菌HTZ1、HTZ2、HTZ3、HTZ4、HTZ5分别被认定为根癌农杆菌(Agrobacterium tumefaciens (Smith et Townsend) Conn)(同源相似性98.16%,下同)、费氏中华根瘤菌(Sinorhizobium fredii (Scholla et Elkan) Chen)(99.82%)、阿拉伯分枝杆菌(Mycolicibacterium arabiense Zhang)(98.88%)、墨西哥假黄单胞菌(Pseudoxanthomonas mexicana Thierry)(97.87%)和日本假黄单胞菌(Pseudoxanthomonas japonensis Thierry)(98.13%)。从天水试验区远根土中另外分离得到的2株菌TS2、TS4,分属纤维弧菌属(Cellvibrio)和固氮菌属(Azotobacter),被认定为Cellvibrio fibrivorans Mergaert(98.17%)和圆褐固氮菌(Azotobacter chroococcum Beijerinck)(99.73%)。从兰州远根土和根表土中另外分离得到的2株菌HP5、HP10分属肠杆菌属(Enterobacter)和鞘氨醇杆菌属(Sphingobium),被认定为烟草肠杆菌(Enterobacter tabaci Duan)(99.53%)和Sphingobium abikonense Kumari(98.70%)。从张掖远根土中分离得的ZY9也属于假黄单胞菌属(Pseudoxanthomonas sp.)(99.38%)。以上菌株在杂交构树根际的分布数量表现为根表土 > 远根土,菌株种类数量最多的为假黄单胞菌属,共3株,优势率为37.5%。3个试验区杂交构树的根内均未分离得到固氮菌。
表 1 杂交构树根际固氮菌分离结果与菌落形态
Table 1. Results and colony morphology of root-bound nitrogen-fixing bacteria from hybrid Broussonetia papyrifera
分离地
Collection sites菌株
Strain分离部位
Separated position菌落直径
Colony dia-meter/mm生长速度
Growth rate菌落形态
Colonial morphology①②③ HTZ1 TS 2.0~3.0 ++ 无色透明,表面凸起,边缘整齐,后期有流动性
Colorless and transparent, convex surface, neat edge, fluidity in later period① TS2 FRS 2.0~3.0 ++ 无色半透明,表面凸起,边缘不规则,后期有流动性
Colorless translucent, convex surface, irregular edge, fluidity in later period①②③ HTZ2 TS 0.5~1.0 ++ 乳白半透明,表面凸起,边缘整齐
Milky white translucent, convex surface, neat edges① TS4 FRS 0.5~1.0 ++ 乳白不透明,表面凹陷,边缘不规则
Opalescent opacity, concave surface, irregular edge② HP5 FRS 1.0~2.0 + 橘黄不透明,表面凸起,边缘整齐,后期生长非常缓慢
Orange and opaque, raised surface, neat edges, very slow growth later①②③ HTZ3 TS 1.0~2.0 +++ 淡黄半透明,表面凸起,有粘性,边缘不规则,后期流动性强
Pale yellow and translucent, convex surface, viscous, irregular edge, strong fluidity in later period①②③ HTZ4 TS 0.5~1.0 +++ 淡黄半透明,表面凸起,有粘性,边缘整齐,后期流动性强
Pale yellow and translucent, convex surface, viscous, neat edge, strong fluidity in later period①②③ HTZ5 TS 0.5~1.0 +++ 淡黄不透明,表面凹陷,有粘性,边缘不规则,后期流动性强
Pale yellow and opaque, sunken surface, viscous, irregular edge, strong fluidity in later period③ ZY9 FRS 0.5~1.0 ++ 乳白不透明,表面凹陷,边缘不规则,后期棕褐
Opalescent opaque, surface sunken, margin irregular, brown in later period② HP10 TS 0.5~1.0 ++ 乳白不透明,表面褶皱,边缘不规则,后期棕褐
Opalescent opacity, surface pleated, margin irregular,brown in later period注:①、②、③分别代表天水、兰州、张掖试验区;分离部位中TS、FRS分别代表根表土、远根土;生长速度中“+、++、+++”分别代表菌株前期生长较慢、中等、较快,48 h内的为生长速度较快,72 h内为中等,96 h内较慢。
Notes: ①, ② and ③ represent Tianshui, Lanzhou and Zhangye experimental areas respectively. TS and FRS represent root topsoil and far rhizosphere soil, respectively."+, ++, +++" respectively represent the slow, medium and fast growth in the early stage of the strain, the fast growth within 48 h, the medium growth within 72 h, and the slow growth within 96 h. -
表2表明:分离得到的10株固氮菌固氮酶活性存在差异,其中,HTZ4的固氮酶活性最低(130.41 IU·L−1),TS4的活性最高(184.51 IU·L−1)。所有固氮菌均无溶解无机磷的能力,仅HTZ2和TS4在蒙金娜有机磷培养上出现溶磷圈,说明这2株菌具有一定溶解有机磷(卵磷脂)的能力(图2、表2),但二者差异不显著,而在发酵液中有机磷的增量表现为TS4显著大于HTZ2。能够分泌IAA的固氮菌有8株,占分离获得菌株数量的80%。HTZ4和HTZ1产IAA显色反应最明显,说明具有较强的分泌IAA的能力,分泌量分别达44.62、36.52 µg·mL−1,与其他菌株差异极显著;HTZ4菌液IAA的分泌量比HP10的高出约20倍(表2)。
表 2 固氮菌固氮酶活性、溶磷和产IAA能力的检测结果
Table 2. Results of activity, dissolved phosphorus and IAA production capacity of nitrogen-fixing bacteria
固氮菌
Nitrogen fixing
bacteria固氮酶活性
Nitrogenase activity/
(IU·L−1)溶解有机磷
Soluble phosphorus
reaction/(mm·mm−1)有机磷增量
Organophosphorus increment/
(µg·mL−1)产IAA显色反应
IAA chromogenic
reactionIAA增量
IAA Increment/
(µg·mL−1)HTZ1 139.53 ± 1.09 E 0.00 ± 0.02 B 0.00 ± 0.12 C +++ 36.52 ± 0.69 B TS2 137.40 ± 2.05 E 0.00 ± 0.05 B 0.00 ± 0.02 C − 0.00 ± 0.00 E HTZ2 163.08 ± 1.06 C 1.18 ± 0.02 A 1.60 ± 0.91 B ++ 8.47 ± 0.29 C TS4 184.51 ± 0.66 A 1.37 ± 0.06 A 3.15 ± 0.31 A + 3.84 ± 0.24 DE HP5 153.05 ± 0.60 D 0.00 ± 0.01 B 0.00 ± 0.22 C + 1.73 ± 0.08 E HTZ3 177.52 ± 0.91 B 0.00 ± 0.02 B 0.00 ± 0.17 C − 0.00 ± 0.09 E HTZ4 130.41 ± 1.61F 0.00 ± 0.05 B 0.00 ± 0.04 C +++ 44.62 ± 0.70 A HTZ5 166.58 ± 1.32 C 0.00 ± 0.11 B 0.00 ± 0.02 C ++ 7.74 ± 0.31 C ZY9 155.64 ± 0.52 D 0.00 ± 0.08 B 0.00 ± 0.00 C + 5.26 ± 0.17 D HP10 182.84 ± 0.55 AB 0.00 ± 0.10 B 0.00 ± 0.00 C + 2.26 ± 0.25 E 注:表中数据为平均值 ± 标准误。同列不同大写字母表示差异极显著(p < 0.01) ;产IAA显色反应中“−、+、+ +、+ + +”分别代表不变色、浅红色、粉红色、红色。
Notes: Data in the table are mean ± standard error. Different capital letters in the same column indicated significant difference (p < 0.01);In IAA reaction, "−, +, + +, + + +" respectively represent non-discoloration, light red, pink, and red.图 2 菌株HTZ2(A)和TS4(B)在蒙金娜有机磷培养基上的解磷能力
Figure 2. Phosphorolysis ability of strains HTZ2(A)and TS4(B)on Montana organophosphate medium
10株固氮菌对不同植物病原菌的拮抗作用不同(表3、图3、4)。所有菌株对Bc均无拮抗作用,除HTZ2、TS4和TS2对另外4种供试病原菌也无抑制效果或效果微弱外,其余7株菌分别对至少2种以上病原菌有一定的抑制作用,其中,HP5、HP10、HTZ4、ZY9对As、Fs、Fo均有拮抗作用,HTZ4、HP5分别对As、Fs的抑菌率最大,分别为50.00%、47.37%,其次为HP10,对As、Fs抑制率分别为45.83%、44.44%;HP5、ZY9对Rs也有拮抗作用。
图 3 HP5菌株与部分病原菌的拮抗试验结果
Figure 3. Antagonistic test results of the HP5 strain against some pathogenic fungi
表 3 固氮菌与植物病原菌的拮抗作用
Table 3. Results of antagonism between nitrogen-fixing bacteria and plant pathogenic fungi
固氮菌
Nitrogen-fixing bacteria抑菌率 Antibacterial rate/% Fs As Fo Bc Rs HTZ1 0.00 ± 0.12 F 18.37 ± 0.02 F 28.57 ± 0.02 C 0.00 0.00 ± 0.11 D TS2 0.00 ± 0.02 F 6.25 ± 0.02 G 0.00 ± 0.11 E 0.00 0.00 ± 0.19 D HTZ2 0.00 ± 0.11 F 0.00 ± 0.10 H 0.00 ± 0.05 E 0.00 0.00 ± 0.10 D TS4 0.00 ± 0.08 F 0.00 ± 0.02 H 0.00 ± 0.13 E 0.00 0.00 ± 0.01 D HP5 47.37 ± 0.76 A 40.00 ± 0.44 C 11.73 ± 0.25 D 0.00 37.58 ± 0.29 B HTZ3 0.00 ± 0.05 F 21.55 ± 0.32 E 48.78 ± 1.01 A 0.00 0.00 ± 0.02 D HTZ4 42.85 ± 0.31 C 50.00 ± 0.54 A 11.76 ± 0.12 D 0.00 0.00 ± 0.04 D HTZ5 25.19 ± 0.45 E 0.00 ± 0.05 H 0.00 ± 0.08 E 0.00 40.74 ± 0.13 A ZY9 27.27 ± 0.18 D 37.50 ± 1.65 D 30.30 ± 0.09 B 0.00 34.61 ± 0.96 C HP10 44.44 ± 0.77 B 45.83 ± 0.76 B 26.82 ± 1.33 C 0.00 0.00 ± 0.05 D 图 4 HP10菌株与部分病原菌的拮抗试验结果
Figure 4. Antagonistic test results of the HP10 strain against some pathogenic fungi
通过对分离得到的10株固氮菌进行固氮、溶磷、分泌生长素、抑菌特性测试结果表明:各菌株对不同测试指标表现各异,其中,TS4、HTZ2 具有固氮、溶磷、分泌IAA的能力,但不具备抑制供试病原菌的能力;HP5、HTZ4、HTZ5、ZY9、HP10除无溶磷能力外,具有不同程度的固氮、分泌IAA、抑制病原菌Fs、As、Fo、Rs的作用,而TS2具有较强的固氮能力。为此,对10株固氮菌的上述4个特性进行了综合评价(表4),排名前5位的菌株为TS4、HP5、ZY9、HTZ4和HTZ5。
表 4 固氮菌特性综合评价结果
Table 4. Comprehensive evaluation results of nitrogen-fixing bacteria characteristics
菌株
Bacterial strain各指标比重值 Specific gravity value of each index 得分
Score排名
Ranking固氮
Nitrogen fixation有机磷增量
Organophosphorus incrementIAA增量
IAA increment抑菌 Bacteriostatic activity Fs As Fo Rs HTZ1 0.031 8 0.000 0 0.330 7 0.000 0 0.083 7 0.180 9 0.000 0 0.078 8 8 TS2 0.024 4 0.000 0 0.000 0 0.000 0 0.028 5 0.000 0 0.000 0 0.004 1 10 HTZ2 0.114 0 0.336 8 0.076 7 0.000 0 0.000 0 0.000 0 0.000 0 0.104 2 6 TS4 0.188 9 0.663 2 0.034 8 0.000 0 0.000 0 0.000 0 0.000 0 0.186 2 1 HP5 0.079 0 0.000 0 0.015 7 0.253 2 0.182 2 0.074 3 0.332 8 0.131 7 2 HTZ3 0.164 5 0.000 0 0.000 0 0.000 0 0.098 2 0.308 8 0.000 0 0.055 5 9 HTZ4 0.000 0 0.000 0 0.404 0 0.229 0 0.227 8 0.074 4 0.000 0 0.118 3 4 HTZ5 0.126 3 0.000 0 0.070 1 0.134 6 0.000 0 0.000 0 0.360 8 0.106 3 5 ZY9 0.088 1 0.000 0 0.047 6 0.145 7 0.170 8 0.191 8 0.306 5 0.130 5 3 HP10 0.183 0 0.000 0 0.020 5 0.237 5 0.208 8 0.169 8 0.000 0 0.084 4 7 熵值 Entropy 0.868 8 0.425 2 0.677 7 0.709 0 0.787 3 0.735 3 0.557 0 权重 Weight 0.058 6 0.256 6 0.143 9 0.129 9 0.095 0 0.118 2 0.197 8
杂交构树根际固氮菌的分离与特性研究
Isolation and Investigation on Properties of Nitrogen-fixing Bacteria from Rhizosphere of Broussonetia papyrifera Hybrid
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摘要:
目的 获得杂交构树根际固氮菌菌株并明确其功能特性。 方法 以Ashby培养基为选择分离条件,从甘肃天水、兰州、张掖种植试验区的杂交构树根系及根际土壤中分离固氮菌,对固氮菌进行鉴定及系统发育分析,同时测定固氮酶活性、溶磷、分泌吲哚乙酸和抑制植物病原菌的能力,并采用熵权法对菌株特性进行综合评价。 结果 从远根土和根表土中共分离出10株固氮菌,分属8个属,以假黄单胞菌属(Pseudoxanthomonas sp.)居多。各菌株均具有固氮酶活性,TS4和HP10的固氮酶活性均大于180 IU·L−1;仅TS4和HTZ2具有溶磷能力;有8株菌能够分泌吲哚乙酸,HTZ4的分泌能力最强,分泌量为44.62 µg·mL−1。所有菌株对灰霉菌(Botrytis cinerea Persoon)无拮抗作用;有7株菌分别对至少2种以上植物病原菌有抑制作用。HTZ4、HTZ3、HP5分别对茄链格孢(Alternaria solani(Ellis et G. Martin) Sorauer)、尖孢镰刀菌(Fusarium oxysporum Schlecht)、腐皮镰孢菌(Fusarium solani (Martius) Sacco)有最强抑制,抑制率分别为50.00%、48.78%、47.37%。综合性能较强的菌株为TS4、HP5、ZY9、HTZ4和HTZ5。 结论 研究结果可为进一步开发综合性能优良的固氮菌株,研制杂交构树多功能固氮菌肥,提高西北半干旱地区杂交构树产量提供依据和指导。 Abstract:Objective To obtain strains of nitrogen-fixing bacteria from Broussonetia papyrifera (Linnaeus) L’Héritier ex Ventenat hybrid and determine their functional characteristics. Method The Ashby medium was selected as separation condition to isolated nitrogen-fixing bacteria from the roots and rhizosphere soil of B. papyrifera hybrid in Tianshui, Lanzhou and Zhangye experimental areas of Gansu Province. In addition, identification and phylogenetic analysis were carried out on the isolated nitrogen-fixing bacteria. The nitrogenase activity, phosphorus solubility, properties of IAA-producing and inhibition of plant pathogenic fungi of each strain were determined at the same time. Entropy weight method was used for comprehensive evaluation of the features of the strains. Result A total of 10 nitrogen-fixing bacteria was isolated from the soil far from the rhizosphere and topsoil, which belonged to 8 genera, mostly Pseudoxanthomonas. Each strain showed nitrogenase activity, among which the nitrogenase activity of the strains TS4 and HP10 was greater than 180 IU·L−1. Only the strains TS4 and HTZ2 had certain phosphorus solubility. 8 strains could secrete IAA, among which the strain HTZ4 demonstrated the strongest secretion ability, with an IAA increment of 44.62 µg·mL−1. All strains showed no antagonistic effect on Botrytis cinerea Persoon, 7 strains showed inhibitory effects on at least two kinds of plant pathogens. The strains HTZ4, HTZ3, and HP5 showed the strongest inhibitory activities against Alternaria solani (Ellis et G. Martin) Sorauer, Fusarium oxysporum Schlecht and Fusarium solani (Martius) Sacco, respectively, with inhibition rates of 50.00%, 48.78% and 47.37%. To sum up, the strains with strong comprehensive properties were the strains TS4, HP5, ZY9, HTZ4 and HTZ5. Conclusion The results of this study can provide data and references for further developing nitrogen-fixing strains with excellent comprehensive performance, multifunctional nitrogen-fixing bacterial fertilizer for Broussonetia papyrifera hybrid, and promoting the yield of Broussonetia papyrifera hybrid in semi-arid area of Northwest China. -
表 1 杂交构树根际固氮菌分离结果与菌落形态
Table 1. Results and colony morphology of root-bound nitrogen-fixing bacteria from hybrid Broussonetia papyrifera
分离地
Collection sites菌株
Strain分离部位
Separated position菌落直径
Colony dia-meter/mm生长速度
Growth rate菌落形态
Colonial morphology①②③ HTZ1 TS 2.0~3.0 ++ 无色透明,表面凸起,边缘整齐,后期有流动性
Colorless and transparent, convex surface, neat edge, fluidity in later period① TS2 FRS 2.0~3.0 ++ 无色半透明,表面凸起,边缘不规则,后期有流动性
Colorless translucent, convex surface, irregular edge, fluidity in later period①②③ HTZ2 TS 0.5~1.0 ++ 乳白半透明,表面凸起,边缘整齐
Milky white translucent, convex surface, neat edges① TS4 FRS 0.5~1.0 ++ 乳白不透明,表面凹陷,边缘不规则
Opalescent opacity, concave surface, irregular edge② HP5 FRS 1.0~2.0 + 橘黄不透明,表面凸起,边缘整齐,后期生长非常缓慢
Orange and opaque, raised surface, neat edges, very slow growth later①②③ HTZ3 TS 1.0~2.0 +++ 淡黄半透明,表面凸起,有粘性,边缘不规则,后期流动性强
Pale yellow and translucent, convex surface, viscous, irregular edge, strong fluidity in later period①②③ HTZ4 TS 0.5~1.0 +++ 淡黄半透明,表面凸起,有粘性,边缘整齐,后期流动性强
Pale yellow and translucent, convex surface, viscous, neat edge, strong fluidity in later period①②③ HTZ5 TS 0.5~1.0 +++ 淡黄不透明,表面凹陷,有粘性,边缘不规则,后期流动性强
Pale yellow and opaque, sunken surface, viscous, irregular edge, strong fluidity in later period③ ZY9 FRS 0.5~1.0 ++ 乳白不透明,表面凹陷,边缘不规则,后期棕褐
Opalescent opaque, surface sunken, margin irregular, brown in later period② HP10 TS 0.5~1.0 ++ 乳白不透明,表面褶皱,边缘不规则,后期棕褐
Opalescent opacity, surface pleated, margin irregular,brown in later period注:①、②、③分别代表天水、兰州、张掖试验区;分离部位中TS、FRS分别代表根表土、远根土;生长速度中“+、++、+++”分别代表菌株前期生长较慢、中等、较快,48 h内的为生长速度较快,72 h内为中等,96 h内较慢。
Notes: ①, ② and ③ represent Tianshui, Lanzhou and Zhangye experimental areas respectively. TS and FRS represent root topsoil and far rhizosphere soil, respectively."+, ++, +++" respectively represent the slow, medium and fast growth in the early stage of the strain, the fast growth within 48 h, the medium growth within 72 h, and the slow growth within 96 h.表 2 固氮菌固氮酶活性、溶磷和产IAA能力的检测结果
Table 2. Results of activity, dissolved phosphorus and IAA production capacity of nitrogen-fixing bacteria
固氮菌
Nitrogen fixing
bacteria固氮酶活性
Nitrogenase activity/
(IU·L−1)溶解有机磷
Soluble phosphorus
reaction/(mm·mm−1)有机磷增量
Organophosphorus increment/
(µg·mL−1)产IAA显色反应
IAA chromogenic
reactionIAA增量
IAA Increment/
(µg·mL−1)HTZ1 139.53 ± 1.09 E 0.00 ± 0.02 B 0.00 ± 0.12 C +++ 36.52 ± 0.69 B TS2 137.40 ± 2.05 E 0.00 ± 0.05 B 0.00 ± 0.02 C − 0.00 ± 0.00 E HTZ2 163.08 ± 1.06 C 1.18 ± 0.02 A 1.60 ± 0.91 B ++ 8.47 ± 0.29 C TS4 184.51 ± 0.66 A 1.37 ± 0.06 A 3.15 ± 0.31 A + 3.84 ± 0.24 DE HP5 153.05 ± 0.60 D 0.00 ± 0.01 B 0.00 ± 0.22 C + 1.73 ± 0.08 E HTZ3 177.52 ± 0.91 B 0.00 ± 0.02 B 0.00 ± 0.17 C − 0.00 ± 0.09 E HTZ4 130.41 ± 1.61F 0.00 ± 0.05 B 0.00 ± 0.04 C +++ 44.62 ± 0.70 A HTZ5 166.58 ± 1.32 C 0.00 ± 0.11 B 0.00 ± 0.02 C ++ 7.74 ± 0.31 C ZY9 155.64 ± 0.52 D 0.00 ± 0.08 B 0.00 ± 0.00 C + 5.26 ± 0.17 D HP10 182.84 ± 0.55 AB 0.00 ± 0.10 B 0.00 ± 0.00 C + 2.26 ± 0.25 E 注:表中数据为平均值 ± 标准误。同列不同大写字母表示差异极显著(p < 0.01) ;产IAA显色反应中“−、+、+ +、+ + +”分别代表不变色、浅红色、粉红色、红色。
Notes: Data in the table are mean ± standard error. Different capital letters in the same column indicated significant difference (p < 0.01);In IAA reaction, "−, +, + +, + + +" respectively represent non-discoloration, light red, pink, and red.表 3 固氮菌与植物病原菌的拮抗作用
Table 3. Results of antagonism between nitrogen-fixing bacteria and plant pathogenic fungi
固氮菌
Nitrogen-fixing bacteria抑菌率 Antibacterial rate/% Fs As Fo Bc Rs HTZ1 0.00 ± 0.12 F 18.37 ± 0.02 F 28.57 ± 0.02 C 0.00 0.00 ± 0.11 D TS2 0.00 ± 0.02 F 6.25 ± 0.02 G 0.00 ± 0.11 E 0.00 0.00 ± 0.19 D HTZ2 0.00 ± 0.11 F 0.00 ± 0.10 H 0.00 ± 0.05 E 0.00 0.00 ± 0.10 D TS4 0.00 ± 0.08 F 0.00 ± 0.02 H 0.00 ± 0.13 E 0.00 0.00 ± 0.01 D HP5 47.37 ± 0.76 A 40.00 ± 0.44 C 11.73 ± 0.25 D 0.00 37.58 ± 0.29 B HTZ3 0.00 ± 0.05 F 21.55 ± 0.32 E 48.78 ± 1.01 A 0.00 0.00 ± 0.02 D HTZ4 42.85 ± 0.31 C 50.00 ± 0.54 A 11.76 ± 0.12 D 0.00 0.00 ± 0.04 D HTZ5 25.19 ± 0.45 E 0.00 ± 0.05 H 0.00 ± 0.08 E 0.00 40.74 ± 0.13 A ZY9 27.27 ± 0.18 D 37.50 ± 1.65 D 30.30 ± 0.09 B 0.00 34.61 ± 0.96 C HP10 44.44 ± 0.77 B 45.83 ± 0.76 B 26.82 ± 1.33 C 0.00 0.00 ± 0.05 D 表 4 固氮菌特性综合评价结果
Table 4. Comprehensive evaluation results of nitrogen-fixing bacteria characteristics
菌株
Bacterial strain各指标比重值 Specific gravity value of each index 得分
Score排名
Ranking固氮
Nitrogen fixation有机磷增量
Organophosphorus incrementIAA增量
IAA increment抑菌 Bacteriostatic activity Fs As Fo Rs HTZ1 0.031 8 0.000 0 0.330 7 0.000 0 0.083 7 0.180 9 0.000 0 0.078 8 8 TS2 0.024 4 0.000 0 0.000 0 0.000 0 0.028 5 0.000 0 0.000 0 0.004 1 10 HTZ2 0.114 0 0.336 8 0.076 7 0.000 0 0.000 0 0.000 0 0.000 0 0.104 2 6 TS4 0.188 9 0.663 2 0.034 8 0.000 0 0.000 0 0.000 0 0.000 0 0.186 2 1 HP5 0.079 0 0.000 0 0.015 7 0.253 2 0.182 2 0.074 3 0.332 8 0.131 7 2 HTZ3 0.164 5 0.000 0 0.000 0 0.000 0 0.098 2 0.308 8 0.000 0 0.055 5 9 HTZ4 0.000 0 0.000 0 0.404 0 0.229 0 0.227 8 0.074 4 0.000 0 0.118 3 4 HTZ5 0.126 3 0.000 0 0.070 1 0.134 6 0.000 0 0.000 0 0.360 8 0.106 3 5 ZY9 0.088 1 0.000 0 0.047 6 0.145 7 0.170 8 0.191 8 0.306 5 0.130 5 3 HP10 0.183 0 0.000 0 0.020 5 0.237 5 0.208 8 0.169 8 0.000 0 0.084 4 7 熵值 Entropy 0.868 8 0.425 2 0.677 7 0.709 0 0.787 3 0.735 3 0.557 0 权重 Weight 0.058 6 0.256 6 0.143 9 0.129 9 0.095 0 0.118 2 0.197 8 -
[1] 刘 冰, 叶建飞, 刘 夙, 等. 中国被子植物科属概览: 依据APG Ⅲ系统[J]. 生物多样性, 2015, 23(2):225-231. [2] 郭 琪, 张 军, 李 鑫, 等. 饲用型杂交构树在甘肃地区的种植适应性研究[J]. 饲料研究, 2020, 43(7):106-110. [3] Peng X J, Liu H, Chen P L, et al. A Chromosome scale genome assembly of Paper mulberry (Broussonetia papyrifera) provides new insights into its forage and papermaking usage[J]. Molecular Plant, 2019, 12(5): 661-677. doi: 10.1016/j.molp.2019.01.021 [4] 狄义宁, 刘鲁峰, 谢林艳, 等. 一株甘蔗内生菌鉴定及其溶磷能力的研究[J]. 作物杂志, 2018, 34(6):68-75. [5] 张 英, 朱 颖, 姚 拓, 等. 分离自牧草根际四株促生菌株(PGPR)互作效应研究[J]. 草业学报, 2013, 22(1):29-37. [6] 杨 豆, 王清海, 万松泽, 等. 2 株毛竹枯梢病拮抗细菌筛选及其促生功能[J]. 林业科学研究, 2020, 33(6):139-147. [7] 傅晓方. 玉米内生固氮菌的分离鉴定及其对小麦幼苗的促生效益研究[D]. 陕西杨凌: 西北农林科技大学, 2011: 11-15. [8] 马 宁, 刘艳霞, 李 想, 等. 基于熵权法综合评价植物根际促生菌对烟草的促生作用[J]. 南京农业大学学报, 2020, 43(5):887-895. [9] 马海宾, 康丽华, 江业根, 等. 联合固氮菌对桉树青枯病菌的抑制作用研究[J]. 林业科学研究, 2007, 20(4):473-476. [10] Madhurankhi G, Suresh D. Plant growth-promoting rhizobacteria-alleviators of abioticstresses in soil: A review[J]. Pedosphere, 2020, 30(1): 40-61. doi: 10.1016/S1002-0160(19)60839-8 [11] 葛安辉, 方 萍, 熊 超, 等. 联合固氮菌叶面接种剂的优化及其在玉米叶际的定殖[J]. 微生物学通报, 2018, 45(6):1303-1313. [12] 毛莲英, 黄 婵, 祝 开, 等. 固氮菌株XD20对不同甘蔗品种的促生效应[J]. 西北植物学报, 2019, 39(1):140-148. [13] 李建宏, 李雪萍, 张建贵, 等. 饲用玉米根际促生菌资源筛选及其特性研究[J]. 草原与草坪, 2017, 37(1):44-49. [14] 马文文, 姚 拓, 蒲小鹏, 等. 东祁连山7种禾草根际溶磷菌筛选及其溶磷特性[J]. 草业科学, 2015, 32(4):515-523. [15] 胡春锦, 史国英, 曾 泉, 等. 两种不同土壤条件下可培养甘蔗内生固氮菌的多样性[J]. 中国土壤与肥料, 2017(2):141-148. [16] 刘彩霞, 焦如珍, 董玉红, 等. 模拟氮沉降对杉木林土壤氮循环相关微生物的影响[J]. 林业科学, 2015, 51(4):96-102. [17] 李 婷, 何 来, 梁泉峰. 非豆科植物的根瘤菌促生机制的研究进展[J]. 中国农业科技导报, 2013, 15(2):97-102. [18] 袁仁文, 刘 琳, 张 蕊, 等. 植物根际分泌物与土壤微生物互作关系的机制研究进展[J]. 中国农学通报, 2020, 36(2):26-35. [19] Hwaseo k H, Hogyun S, Kyung-Jin K. Structural insights into a maleylpyruvate hydrolase from Sphingobium sp. SYK-6, a bacterium degrading lignin-derived aryls[J]. Biochemical and Biophysical Research Communications, 2019, 514(3): 765-771. doi: 10.1016/j.bbrc.2019.05.030 [20] Hamdy A H, Somya E D, Mostafa M E S. Monitoring the degradation capability of novel haloalkaliphilic tributyltin chloride (TBTCl) resistant bacteria from butyltin-polluted site[J]. Revista Argentina de Microbiología, 2019, 51(1): 39-46. [21] Zhou Y M, Chen Y P, Guo J S, et al. The correlations and spatial characteristics of microbiome and silage quality by reusing of citrus waste in a family-scale bunker silo[J]. Journal of Cleaner Production, 2019, 226: 407-418. doi: 10.1016/j.jclepro.2019.04.075 [22] 王金山, 刘金升, 彭献军, 等. 杂交构树在滨海盐碱地生态绿化中的应用[J]. 天津农业科学, 2014, 20(2):95-101.