林业科学 ›› 2022, Vol. 58 ›› Issue (12): 21-31.doi: 10.11707/j.1001-7488.20221203
曹俐,王阳,杨蕴力,郑雨,王伟,刘桂丰,姜静*
收稿日期:
2022-03-10
出版日期:
2022-12-25
发布日期:
2023-03-11
通讯作者:
姜静
Li Cao,Yang Wang,Yunli Yang,Yu Zheng,Wei Wang,Guifeng Liu,Jing Jiang*
Received:
2022-03-10
Online:
2022-12-25
Published:
2023-03-11
Contact:
Jing Jiang
摘要:
目的: 通过研究转BpGLK裂叶桦根际土壤理化性质、酶活性及微生物群落组成特征,为后续转BpGLK裂叶桦的环境安全性评价及产业化提供理论依据。方法: 以2年生的转BpGLK裂叶桦(RE1~RE5)为研究对象,检测外源基因稳定性;测定其生长特性、根际土壤理化性质及土壤酶活性;利用高通量测序技术分析根际土壤微生物群落组成特征。结果: 转BpGLK株系中的外源Bar基因及BpGLK干扰片段仍然整合于其基因组中,在生长方面,RE株系高于或显著高于WT株系(RE3例外)。RE株系的土壤pH值、有机碳、全氮、速效氮含量低于或显著低于WT株系;而速效磷、全钾及速效钾含量显著高于WT株系(RE1例外)(P < 0.05);RE株系的土壤脲酶、蔗糖酶活性均低于或显著低于WT株系,而中性蛋白酶活性则显著高于WT株系。微生物群落丰富度指数及多样性指数分析显示,在细菌方面:RE株系与WT株系间无显著差异;而真菌的丰富度高于或显著高于WT株系,多样性则与WT株系差异不显著;RE株系的Saccharimonadales、酸热菌属等8个菌属显著高于WT株系,而Candidatus_Solibacter等4个菌属则显著低于WT株系;真菌群落中棉革菌属是RE株系根际土壤的优势类群。结论: 2年生的转BpGLK裂叶桦生长表现优于野生型,尚未发现转基因活动对根际土壤理化性质、土壤酶活性及微生物群落组成产生不利影响。
中图分类号:
曹俐,王阳,杨蕴力,郑雨,王伟,刘桂丰,姜静. 转BpGLK裂叶桦生长变异、根际土壤酶活性及微生物群落组成[J]. 林业科学, 2022, 58(12): 21-31.
Li Cao,Yang Wang,Yunli Yang,Yu Zheng,Wei Wang,Guifeng Liu,Jing Jiang. Growth Variation, Rhizosphere Soil Enzyme Activity and Microbial Community Composition of Transgenic BpGLK Betula pendula 'Dplecprlicp'[J]. Scientia Silvae Sinicae, 2022, 58(12): 21-31.
表1
PCR扩增引物序列"
引物名称 Primer name | 引物序列(5′-3′) Primer sequence | 扩增片段长度 Amplified fragment length/bp |
BpGLK_RNAi_Cis_F | CATGCCATGGGCACAGAAGGTTTGTGCAAG | 414 |
BpGLK_RNAi_Cis_R | TTGGCGCGCCCCATACATCTGCCTTCTCTGG | |
BpGLK_RNAi_Anti_F | GCTCTAGAGCACAGAAGGTTTGTGCAAG | 429 |
BpGLK_RNAi_Anti_R | CGCGGATCCCCATACATCTGCCTTCTCTGG | |
Bar-F | TTAGATCTCGGTGACGGGCA | 500 |
Bar-R | CGGTCTGCACCATCGTCAAC |
表2
根际土壤理化性质比较①"
株系 Strain | pH值 pH value | 有机碳 Organic carbon/(g·kg-1) | 全氮 Total nitrogen/(g·kg-1) | 全磷 Total phosphorus/(g·kg-1) | 全钾 Total potassium/(g·kg-1) | 速效氮 Available nitrogen/(mg·kg-1) | 速效磷 Available phosphorus/(mg·kg-1) | 速效钾 Available potassium/(mg·kg-1) |
WT | 5.06±0.05a | 76.80±1.15a | 4.05±0.06a | 0.73±0.08a | 22.10±0.06c | 364.70±3.70a | 26.30±4.91c | 99.67±0.88b |
RE1 | 4.93±0.09ab | 67.83±0.69c | 3.70±0.14b | 0.66±0.03a | 24.40±0.55b | 338.63±10.34b | 58.50±3.89b | 79.33±8.41c |
RE2 | 4.70±0.12b | 66.90±0.15c | 3.69±0.07b | 0.72±0.08a | 26.23±0.67a | 318.00±0.23c | 82.20±8.66a | 122.67±3.76a |
RE3 | 4.65±0.08b | 70.77±0.97b | 3.73±0.04b | 0.72±0.01a | 24.37±0.20b | 361.35±1.36a | 83.50±4.04a | 111.67±6.89ab |
表3
参试株系根际土壤酶活性比较"
株系 Strain | 脲酶活性 Urease activity/(μg·d-1g-1) | 蔗糖酶活性 Invertase activity/(mg·d-1g-1) | 中性蛋白酶活性 Neutral protease activity/(mg·d-1g-1) |
WT | 658.73±17.66a | 32.38±1.09a | 5.29±0.04c |
RE1 | 455.97±12.24b | 11.63±0.74c | 7.65±0.48a |
RE2 | 459.46±18.35b | 29.92±0.42a | 6.38±0.19b |
RE3 | 433.83±14.02b | 18.75±1.70b | 7.07±0.40ab |
表4
根际土壤细菌及真菌α多样性分析"
微生物 Microbial | 样品 Samples | 检测到的物种数 Observed_species | Chao1指数 Chao1 index | 香农指数 Shannon index | Simpson指数 Simpson index | 测序深度指数 Coverage |
细菌 Bacteria | WT | 6 217.03±192.10a | 7 221.13±307.51a | 11.20±0.07a | 0.999±0.000a | 0.944±0.004a |
RE1 | 5 705.98±324.68a | 6 862.75±589.74a | 10.93±0.09a | 0.998±0.000a | 0.945±0.007a | |
RE2 | 6 091.20±461.23a | 7 081.53±634.46a | 11.09±0.19a | 0.998±0.000a | 0.945±0.007a | |
RE3 | 5 503.93±665.56a | 6 459.25±931.47a | 10.86±0.26a | 0.998±0.000a | 0.950±0.010a | |
真菌 Fungi | WT | 206.95±19.59b | 208.45±20.15b | 3.22±0.28ab | 0.790±0.044a | 0.999±0.000a |
RE1 | 339.10±46.95a | 341.53±46.70a | 4.00±0.29a | 0.841±0.028a | 0.998±0.000a | |
RE2 | 250.50±17.37ab | 252.75±17.42ab | 3.17±0.25b | 0.748±0.036a | 0.998±0.000a | |
RE3 | 209.43±22.64b | 211.02±23.05b | 3.39±0.12ab | 0.827±0.026a | 0.998±0.000a |
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