Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (12): 223-232.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0078
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DU Jia-hui1(), XU Wei-fang1(), YANG Xiao-dong1, TAN Song1, YIN Deng-ke1,2,3, LIU Yuan-xu1,2()
Received:
2022-01-19
Online:
2022-12-26
Published:
2022-12-29
Contact:
XU Wei-fang,LIU Yuan-xu
E-mail:1650268474@qq.com;xuweifang@ahtcm.edu.cn;yuanxliu@ahtcm.edu.cn
DU Jia-hui, XU Wei-fang, YANG Xiao-dong, TAN Song, YIN Deng-ke, LIU Yuan-xu. Isolation and Screening of Endophytes Producing Indole Acetic Acid from Polygonatum cyrtonema Hua. and Its Effect on Seed Germination of Polygonatum[J]. Biotechnology Bulletin, 2022, 38(12): 223-232.
培养基 Medium | 内生细菌 Endophytic bacteria | 内生真菌 Endophytic fungi |
---|---|---|
PDA培养基 PDA medium | 10 | 7 |
LB培养基 LB medium | 6 | 0 |
NA培养基NA medium | 8 | 0 |
GA培养基GA medium | 1 | 1 |
CZA培养基 CZA medium | 0 | 1 |
Table 1 Statistics of endophytes isolated from Polygonatum cyrtonema Hua.
培养基 Medium | 内生细菌 Endophytic bacteria | 内生真菌 Endophytic fungi |
---|---|---|
PDA培养基 PDA medium | 10 | 7 |
LB培养基 LB medium | 6 | 0 |
NA培养基NA medium | 8 | 0 |
GA培养基GA medium | 1 | 1 |
CZA培养基 CZA medium | 0 | 1 |
Fig. 2 Morphology of endophytic fungi isolated from Pol-ygonatum cyrtonema Hua. on PDA medium Left:Colony morphology on the front plate;Right:colony morphology on the reverse plate
菌株编号 Strain No. | 产IAA强度 IAA production strength |
---|---|
IAA标准品 IAA standard substance(30 mg·L-1) | +++ |
HNC2、HPB1 | ++ |
HPB3、HPC2、HPC4、HPA3、HLB3 | + |
HLB2、HNC1、HNB3、HGC1、HPB2、HNC3、HLA3、HPA2、HNA3、HPA1、HLA1、HLA4、HNB1、HNB2、HNA2、HPC3、HPC1、HLA2 | - |
Table 2 Statistics of IAA-producing ability of endophytic bacteria isolated from Polygonatum cyrtonema Hua.
菌株编号 Strain No. | 产IAA强度 IAA production strength |
---|---|
IAA标准品 IAA standard substance(30 mg·L-1) | +++ |
HNC2、HPB1 | ++ |
HPB3、HPC2、HPC4、HPA3、HLB3 | + |
HLB2、HNC1、HNB3、HGC1、HPB2、HNC3、HLA3、HPA2、HNA3、HPA1、HLA1、HLA4、HNB1、HNB2、HNA2、HPC3、HPC1、HLA2 | - |
Fig. 4 IAA standard curve(A)and trends of the content of IAA secreted by endophytic bacteria of Polygona-tum cyrtonema Hua. over time(B) The error line in the figure refers to the standard deviation. Different lowercase letters represent significant differences at 0.05 level
Fig. 5 Colony and morphological characteristics of HNC2 strain and HPB1 strain A:Colony morphology of HNC2 strain. B:Gram stain result of HNC2 strain. C:Spore stain result of HNC2 strain. D:Colony morphology of HPB1 strain. E:Gram stain result of HPB1 strain
生理生化测试项目 Test item | 结果 Results | ||
---|---|---|---|
HNC2 | HPB1 | ||
葡萄糖 Glucose | + | - | |
乳糖 Lactose | - | - | |
麦芽糖 Maltose | + | - | |
甘露醇 Mannitol | - | - | |
蔗糖 Sucrose | + | - | |
靛基质试验 Indole test | + | + | |
甲基红试验Methyl Red(MR) | + | - | |
乙酰甲基甲醇试验 Voges-Proskaurer(VP) | + | + | |
西蒙氏柠檬酸盐 Simon’s citrate | + | + | |
硫化氢 Hydrogen sulphide | - | - | |
尿素 Urea | - | + | |
半固体琼脂 Semi-solid agar | + | + |
Table 3 Physiological and biochemical characteristics of HNC2 strain and HPB1 strain
生理生化测试项目 Test item | 结果 Results | ||
---|---|---|---|
HNC2 | HPB1 | ||
葡萄糖 Glucose | + | - | |
乳糖 Lactose | - | - | |
麦芽糖 Maltose | + | - | |
甘露醇 Mannitol | - | - | |
蔗糖 Sucrose | + | - | |
靛基质试验 Indole test | + | + | |
甲基红试验Methyl Red(MR) | + | - | |
乙酰甲基甲醇试验 Voges-Proskaurer(VP) | + | + | |
西蒙氏柠檬酸盐 Simon’s citrate | + | + | |
硫化氢 Hydrogen sulphide | - | - | |
尿素 Urea | - | + | |
半固体琼脂 Semi-solid agar | + | + |
Fig. 6 Phylogenetic tree of HNC2 strain(A)and HPB1 strain(B)based on 16S rDNA sequence Number in parentheses is the GenBank accession numbers. Number at the nodes indicates the Bootstrap value. The scale bar indicates 0.5% and 2% nucleotide substitution,respectively
处理方法 Treatment | 发芽势 Germination potential/% | 发芽率 Germination rate/% | 胚芽长 Germ length/mm | 胚根长 Radicle length/mm | 叶宽 Leaf width/mm |
---|---|---|---|---|---|
无菌水Sterile water | 6.67±1.67 cd | 11.67±1.67 cd | 19.47±3.73 | 6.83±1.15 b | 1.73±0.39 |
LB液体培养基LB liquid medium | 5.00±0.00 d | 10.00±2.89 d | 22.73±5.25 | 14.13±2.72 ab | 2.83±0.12 |
IAA标准品IAA standard substance | 15.00±0.00 abc | 21.67±4.41 abc | 21.17±2.81 | 21.13±7.09 a | 2.73±1.23 |
HNC2发酵液HNC2 supernatant | 23.33±1.67 a | 26.67±1.67 a | 19.70±0.74 | 14.43±0.93 ab | 2.23±0.12 |
HNC2菌悬液HNC2 cell suspension | 18.33±3.33 ab | 25.00±5.77 ab | 20.10±1.19 | 14.60±1.85 ab | 2.73±0.42 |
HPB1发酵液HPB1 supernatant | 11.67±1.67 bcd | 15.00±2.89 bcd | 16.80±3.02 | 15.10±2.66 ab | 3.03±1.80 |
HPB1菌悬液HPB1 cell suspension | 21.67±6.01 a | 28.33±1.67 a | 18.20±0.61 | 12.23±0.79 ab | 2.53±0.23 |
Table 4 Effects of endophytic bacteria HNC2 and HPB1 on the germination of Polygonatum seeds
处理方法 Treatment | 发芽势 Germination potential/% | 发芽率 Germination rate/% | 胚芽长 Germ length/mm | 胚根长 Radicle length/mm | 叶宽 Leaf width/mm |
---|---|---|---|---|---|
无菌水Sterile water | 6.67±1.67 cd | 11.67±1.67 cd | 19.47±3.73 | 6.83±1.15 b | 1.73±0.39 |
LB液体培养基LB liquid medium | 5.00±0.00 d | 10.00±2.89 d | 22.73±5.25 | 14.13±2.72 ab | 2.83±0.12 |
IAA标准品IAA standard substance | 15.00±0.00 abc | 21.67±4.41 abc | 21.17±2.81 | 21.13±7.09 a | 2.73±1.23 |
HNC2发酵液HNC2 supernatant | 23.33±1.67 a | 26.67±1.67 a | 19.70±0.74 | 14.43±0.93 ab | 2.23±0.12 |
HNC2菌悬液HNC2 cell suspension | 18.33±3.33 ab | 25.00±5.77 ab | 20.10±1.19 | 14.60±1.85 ab | 2.73±0.42 |
HPB1发酵液HPB1 supernatant | 11.67±1.67 bcd | 15.00±2.89 bcd | 16.80±3.02 | 15.10±2.66 ab | 3.03±1.80 |
HPB1菌悬液HPB1 cell suspension | 21.67±6.01 a | 28.33±1.67 a | 18.20±0.61 | 12.23±0.79 ab | 2.53±0.23 |
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