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作物学报 ›› 2022, Vol. 48 ›› Issue (4): 860-872.doi: 10.3724/SP.J.1006.2022.14052

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

不同施肥水平下丛枝菌根真菌对甘蔗生长及养分相关基因共表达网络的影响

孔垂豹1(), 庞孜钦1,2, 张才芳1, 刘强1,2, 胡朝华1, 肖以杰1, 袁照年1,3,*()   

  1. 1福建农林大学国家甘蔗工程技术研究中心, 福建福州 350002
    2福建农林大学农学院, 福建福州 350002
    3蔗糖产业省部共建协同创新中心, 广西南宁 530000
  • 收稿日期:2021-03-31 接受日期:2021-07-12 出版日期:2022-04-12 网络出版日期:2021-07-26
  • 通讯作者: 袁照年
  • 作者简介:E-mail: kongchuibao18@163.com
  • 基金资助:
    国家现代农业产业技术体系建设专项(CARS-170208);福建农林大学科技创新项目资助(KFA17528A)

Effects of arbuscular mycorrhizal fungi on sugarcane growth and nutrient- related gene co-expression network under different fertilization levels

KONG Chui-Bao1(), PANG Zi-Qin1,2, ZHANG Cai-Fang1, LIU Qiang1,2, HU Chao-Hua1, XIAO Yi-Jie1, YUAN Zhao-Nian1,3,*()   

  1. 1National Engineering Research Center for Sugarcane, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
    2College of Agriculture, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China
    3Province and Ministry Co-sponsored Collaborative Innovation Center of Sugar Industry, Nanning 530000, Guangxi, China
  • Received:2021-03-31 Accepted:2021-07-12 Published:2022-04-12 Published online:2021-07-26
  • Contact: YUAN Zhao-Nian
  • Supported by:
    China Agriculture Research System(CARS-170208);Science and Technology Innovation Project of Fujian Agriculture and Forestry University(KFA17528A)

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摘要：

甘蔗是我国重要的糖料作物之一。丛枝菌根(arbuscular mycorrhizas, AM)真菌在自然界中分布广泛, 研究表明AM真菌侵染植物根系能够促进植物的养分吸收和生长发育。本研究利用盆栽试验设置常规施肥(N)和减量施肥(R) 2个施肥水平, 并分别进行接种(AM)和未接种(CK) AM真菌处理, 共计4组, 每组设置4个重复。结果表明, 接种AM真菌不仅可以显著提高甘蔗地上及地下部生物量的积累, 而且还显著影响甘蔗根际土壤pH值、碱解氮、有效磷的含量。同时, 接种AM真菌在减量施肥水平下对甘蔗生物量的提高作用明显高于常规施肥水平。通过加权基因共表达网络分析(weighted gene co-expression network analysis, WGCNA)筛选到与氮、磷等养分表型存在高度特异性的turquoise模块和darkgreen模块。以KME值大于0.7为阈值筛选模块的核心基因, 分别筛选到408个和21个核心基因。通过GO富集分析发现, 这些核心基因主要参与养分转运、代谢以及催化活性等通路。根据基因的注释信息以及基因间的连通性, 在筛选出的核心基因中寻找到28个与氮、磷等养分吸收与转运相关的核心基因以及108个候选基因。本研究揭示了AM真菌对甘蔗养分吸收的影响效应, 为进一步理解AM真菌影响甘蔗养分吸收的分子机制提供了理论基础。

关键词: 丛枝菌根真菌, 甘蔗生长, 养分, 加权基因共表达网络

Abstract:

Sugarcane is one of the important sugar crops in China. Arbuscular mycorrhizas (AM) fungi are widely distributed. Researches have shown that AM fungi infecting plant roots can promote nutrient absorption and growth of plants. In this study, pot experiment was used to set up with two fertilization levels of conventional fertilization (N) and reduced fertilization (R), and inoculated (AM) and control (CK). There were four treatments in total, and four replicates were set in each treatment. The results revealed that the plants inoculated with AM fungi did not only significantly increased the biomass accumulation, but also significantly affected the pH value, alkali hydrolyzable nitrogen and available phosphorus of sugarcane rhizosphere soil. The biomass accumulation in the AM fungus inoculation treatment under reduced fertilization was significantly higher than that of conventional fertilization. The turquoise module and darkgreen module with high specificity with nutrient phenotypes such as nitrogen and phosphorus were screened by weighted gene co-expression network analysis (WGCNA). The core genes of the module were screened with KME value greater than 0.7 as the threshold, and 408 and 21 core genes were screened, respectively. GO enrichment indicated that these core genes were mainly involved in nutrient transport, metabolism, and enzyme catalysis pathways. Based on annotation information and the connectivity of genes, 28 core genes related to the absorption and transportation of nutrients such as nitrogen and phosphorus and 108 related candidate genes were detected among the core genes screened. This study reveals the effects of AM fungi on sugarcane nutrient absorption, and provides a theoretical basis for further understanding of the molecular mechanisms of AM fungi affecting sugarcane nutrient absorption.

Key words: arbuscular mycorrhizal fungi, sugarcane growth, nutrients, WGCNA

表1

不同处理对甘蔗植株地上部生长指标及菌丝侵染率的影响"

处理
Treatment		 植株干重
Plant biomass (g)		 株高
Plant height (cm)		 菌丝侵染率
Mycelial infection rate (%)
AMN 31.00±0.20 a 41.43±3.27 a 31.25±4.50 b
AMR 27.47±0.84 b 40.09±4.87 a 50.25±8.14 a
CKN 26.37±0.59 b 29.93±0.92 b 0.25±0.25 c
CKR 21.24±0.24 c 23.46±1.13 b 0.50±0.29 c

图1

AM真菌接种处理的真菌产孢及根部菌丝侵染图像 A: 接种AM真菌处理的产孢图像; B: 接种AM真菌处理的甘蔗根部菌丝侵染图像。AM: 丛枝菌根。"

表2

不同处理对甘蔗根干重、根长及根表面积的影响"

处理
Treatment		 根干重
Root biomass (g)		 根长
Length (cm)		 根表面积
Surface area (cm2)
AMN 2.85±0.12 ab 15055.92±162.27 b 1636.08±54.34 a
AMR 3.13±0.06 a 17668.88±220.10 a 1676.34±20.91 a
CKN 2.56±0.02 b 12742.93±158.71 c 1258.49±65.41 c
CKR 2.71±0.20 b 13360.82±365.33 c 1408.73±23.45 b

表3

不同处理对甘蔗根际土壤理化性质的影响"

处理
Treatment		 酸碱度
pH		 碱解氮
AN (mg kg-1)		 有效磷
AP (mg kg-1)		 速效钾
AK (mg kg-1)		 有机质
OM (g kg-1)
AMN 6.75±0.02 b 42.47±2.52 a 12.84±0.25 a 27.38±1.28 a 9.82±0.25 a
AMR 6.92±0.03 a 36.95±3.48 a 10.86±0.37 b 25.63±0.13 ab 7.28±0.14 b
CKN 6.68±0.02 c 26.99±1.52 b 8.89±0.50 c 25.13±1.01 ab 6.79±0.22 b
CKR 6.73±0.02 bc 24.66±1.19 b 7.55±0.16 d 24.00±0.35 b 4.14±0.02 c

图2

软阈值的确定 A的纵坐标表示无尺度网络模型指数; B的纵坐标表示每一个软阈值对应的网络平均连接度; A和B的横坐标均表示软阈值(β)。"

图3

基因聚类树及模块划分 A: 基于拓扑重叠构建的基因聚类树; B: 使用动态剪切算法得到的基因模块, 颜色代表模块; C: 合并相似表达模式后的基因模块。"

图4

模块与性状相关性热图 横坐标表示不同的性状, 纵坐标表示每一个模块的特征向量。方格内的数据表示模块与性状之间的相关性及P值。红色代表模块与性状正相关, 蓝色代表模块与性状负相关。"

表4

模块核心基因GO富集情况(部分)"

模块
Module		 GO富集项目
GO term		 GO 基因数目
Number of genes		 P值
P-value
碧绿Turquoise 代谢过程Metabolic process GO:0008152 122 2.40E-04
细胞过程Cellular process GO:0009987 119 4.70E-03
单一生物过程Single-organism process GO:0044699 99 4.13E-03
生物调节Biological regulation GO:0065007 37 4.27E-03
对刺激的反应Response to stimulus GO:0050896 35 4.78E-03
本质化过程Localization GO:0051179 29 4.74E-03
细胞Cell GO:0005623 133 5.11E-04
细胞部分Cell part GO:0044464 133 2.39E-03
细胞器Organelle GO:0043226 100 7.90E-05
膜Membrane GO:0016020 92 2.04E-03
膜部分Membrane part GO:0044425 75 2.52E-03
细胞器部分Organelle part GO:0044422 29 3.51E-03
结合Binding GO:0005488 108 2.58E-03
催化活性Catalytic activity GO:0003824 98 7.54E-03
深绿Darkgreen 代谢过程Metabolic process GO:0008152 13 1.25E-03
细胞过程Cellular process GO:0009987 8 3.45E-03
单一生物过程Single-organism process GO:0044699 8 2.54E-03
膜Membrane GO:0016020 8 3.25E-03
细胞Cell GO:0005623 7 6.45E-03
细胞部分Cell part GO:0044464 7 5.87E-03
膜部分Membrane part GO:0044425 7 2.58E-03
细胞器Organelle GO:0043226 6 4.65E-03
催化活性Catalytic activity GO:0003824 11 2.35E-03
结合Binding GO:0005488 8 4.78E-03

表5

各基因模块中养分相关核心基因功能注释(部分)"

模块
Module		 基因ID
Gene ID		 基因功能
Gene function
碧绿 Saccharum_officinarum_newGene_73305 蔗糖合酶2
Turquoise Sucrose synthase 2
Saccharum_officinarum_newGene_56251 氨基酸转运和代谢
Amino acid transport and metabolism
Saccharum_officinarum_newGene_107291 氨基酸转运和代谢
Amino acid transport and metabolism
Saccharum_officinarum_newGene_109658 可溶性无机焦磷酸酶; 无机离子迁移与代谢
Soluble inorganic pyrophosphatase; inorganic ion transport and metabolism
Saccharum_officinarum_newGene_84225 碳水化合物的运输和代谢
Carbohydrate transport and metabolism
Saccharum_officinarum_newGene_20999 氨基酸转运和代谢
Amino acid transport and metabolism
Saccharum_officinarum_newGene_56328 碳水化合物的运输和代谢
Carbohydrate transport and metabolism
模块
Module		 基因ID
Gene ID		 基因功能
Gene function
Saccharum_officinarum_newGene_61584 氨基酸转运和代谢
Amino acid transport and metabolism
Saccharum_officinarum_newGene_20996 氨基酸转运和代谢
Amino acid transport and metabolism
Saccharum_officinarum_newGene_36202 钾离子通道基因CNGC2
Cyclic nucleotide-gated ion channel 2
Saccharum_officinarum_newGene_101569 可溶性无机焦磷酸酶
Soluble inorganic pyrophosphatase
Saccharum_officinarum_newGene_36232 钾离子通道基因CNGC2
Cyclic nucleotide-gated ion channel 2
Saccharum_officinarum_newGene_60132 碳水化合物的运输和代谢
Carbohydrate transport and metabolism
Saccharum_officinarum_newGene_116537 主要诱导超级家族; 糖(及其他)转运蛋白
Major Facilitator Super family; sugar (and other) transporter
Saccharum_officinarum_newGene_68404 淀粉和蔗糖代谢; 氨基糖和核苷酸糖代谢
Starch and sucrose metabolism; amino sugar and nucleotide sugar metabolism
Saccharum_officinarum_newGene_52856 碳水化合物的运输和代谢
Carbohydrate transport and metabolism
Saccharum_officinarum_newGene_61188 主要诱导超级家族; 糖(及其他)转运蛋白
Major Facilitator Super family; sugar (and other) transporter
Saccharum_officinarum_newGene_60391 氨基酸转运和代谢
Amino acid transport and metabolism
Saccharum_officinarum_newGene_54480 主要诱导超级家族; 糖(及其他)转运蛋白
Major Facilitator Super family; sugar (and other) transporter
Saccharum_officinarum_newGene_116260 NRT1/PTR FAMILY 7.2蛋白基因
Protein NRT1/PTR FAMILY 7.2
Saccharum_officinarum_newGene_52960 氨基酸转运和代谢
Amino acid transport and metabolism
Saccharum_officinarum_newGene_76759 多肌醇多磷酸磷酸酶1
Multiple inositol polyphosphate phosphatase 1
Saccharum_officinarum_newGene_113779 NRT1/PTR FAMILY 6.4蛋白基因
Protein NRT1/PTR FAMILY 6.4
Saccharum_officinarum_newGene_1327 NRT1/PTR FAMILY 6.4蛋白基因
Protein NRT1/PTR FAMILY 6.4
深绿 Saccharum_officinarum_newGene_108443 碳水化合物的运输和代谢
Darkgreen Carbohydrate transport and metabolism
Saccharum_officinarum_newGene_105049 磷酸酯酶家族基因
Phosphoesterase family
Saccharum_officinarum_newGene_48314 可溶性无机焦磷酸酶; 能源生产与转化
Soluble inorganic pyrophosphatase; energy production and conversion
Saccharum_officinarum_newGene_90724 碳水化合物的运输和代谢
Carbohydrate transport and metabolism

图5

turquoise模块中养分相关核心基因共表达网络 点的大小及颜色的深浅代表点在网络中连通度的高低; 线的颜色深浅代表两点之间连通度的高低。"

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