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作物学报 ›› 2019, Vol. 45 ›› Issue (8): 1189-1199.doi: 10.3724/SP.J.1006.2019.82058

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

不同氮水平下水稻钾吸收及全基因组关联分析

邹伟伟1,路雪丽2,王丽1,薛大伟3,曾大力2,*(),李志新1,*()   

  1. 1 长江大学农学院, 湖北荆州 434025
    2 中国水稻研究所, 浙江杭州 310006
    3 杭州师范大学生命科学学院, 浙江杭州311121
  • 收稿日期:2018-11-22 接受日期:2019-01-19 出版日期:2019-08-12 网络出版日期:2019-03-11
  • 通讯作者: 曾大力,李志新
  • 作者简介:E-mail: 1058510926@qq.com
  • 基金资助:
    本研究由主要粮食作物产业化湖北省协同创新中心资助(MS2015004)

Potassium uptake and genome-wide association analysis of rice under different nitrogen levels

ZOU Wei-Wei1,LU Xue-Li2,WANG Li1,XUE Da-Wei3,ZENG Da-Li2,*(),LI Zhi-Xin1,*()   

  1. 1 Agronomy Department, Yangtze University, Jingzhou 434025, Hubei, China
    2 China National Rice Research Institute, Hangzhou 310006, Zhejiang, China
    3 School of Life Sciences, Hangzhou Normal University, Hangzhou 311121, Zhejiang, China
  • Received:2018-11-22 Accepted:2019-01-19 Published:2019-08-12 Published online:2019-03-11
  • Contact: Da-Li ZENG,Zhi-Xin LI
  • Supported by:
    The study was supported by the Hubei Collaborative Innovation Center for Industrialization of Major Grain Crops(MS2015004)

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摘要：

以业已完成深度重测序的134份水稻地方种质资源为材料, 在大田栽培条件下按不施氮(N0), 施96 kg hm -2纯氮(N1)和施192 kg hm -2纯氮(N2) 3种氮肥水平, 检测了分蘖盛期植株的钾含量、植株干重和钾积累。结果表明, 水稻钾含量、植株干重和钾积累在N0、N1、N2三种氮肥处理下均呈正态分布, 表型变异丰富。植株干重和钾积累与施氮量呈极显著正相关, 钾含量与施氮量相关性不显著; 钾含量与植株干重呈负相关, 钾含量与钾积累相关性不显著, 而植株干重与钾积累呈极显著正相关。在3个施氮水平下, 籼稻的钾含量极显著低于粳稻, 籼稻的干重和钾积累都极显著高于粳稻。全基因组关联分析发现, 在3个氮肥水平下检测到12个显著相关位点, 其中钾积累、钾含量和植株干重的显著相关位点分别有2、5和5个。在N1水平下, 位于第6染色体上与钾含量相关的SNP (Chr6_1,524,776)的显著性峰候选区包含与钾离子转运蛋白互作的RUPO基因。根据钾含量的差异, 鉴定出3个钾含量与低氮响应有关的SNP位点, 1个位点与高氮响应有关, 而位于第10染色体上的显著性位点Chr10_2,822,026对低氮和高氮均有响应, 该区域的4个候选基因的表达在不同氮水平间存在差异。

关键词: 全基因组关联分析, 钾含量, 干重, 钾积累, 水稻

Abstract:

A total of 134 resequenced rice landraces were used for assessing the potassium content, plant dry weight and potassium accumulation at three different nitrogen levels including no nitrogen fertilizer (N0), 96 kg ha -1 (N1), and 192 kg ha -1 (N2) under normal field cultivation, respectively. All the three traits displayed normal distribution with abundant variations under N0, N1, and N2 nitrogen levels, respectively. K accumulation and plant dry weight showed positive correlation with nitrogen levels. Meanwhile, the negative correlation was detected between K content and dry weight, and there was positive correlation between dry weight and K accumulation. In addition, the K content showed significantly lower in indica than in japonica, while the dry weight and K accumulation in indica were significantly higher than those in japonica. A total of 12 SNPs presented significant association with the potassium content, plant dry weight and potassium accumulation under three diferent nitrogen levels, including two SNPs for K accumulation, five SNPs for K content and five SNPs for dry weight. A SNP (Chr6_1,524,776) associated with potassium content on chromosome 6 was detected at N1 level. Its flank contained a receptor-like kinase, RUPO, which interacts with potassium transporters in rice. According to the difference of potassium content, one SNP and three SNPs were identified with high nitrogen and low nitrogen response, respectively. While four candidate genes closed to the SNP (Chr10_2,822,026) were associated to K content relatively changed under both high nitrogen and low nitrogen levels, showing different expression levels under different nitrogen levels.

Key words: genome-wide association study, potassium content, dry weight, potassium accumulation, Oryza sativa

表1

RT-PCR引物"

候选基因
Candidate gene		 上游引物序列
Forward primer sequences (5°-3°)		 下游引物序列
Reverse primer sequences (5'-3')
LOC_Os10g05730 GGCATCAAAACAGGCTCACA GGCCAACTTTACAGGTACACA
LOC_Os10g05600 GCTGTTCTTGGTCCACGC GGCCTTACTGAGTCTCTCCC
LOC_Os10g05540 CTCGGCTTCTTCCACAACTC CCTGGATCTGGCTGGAGAC
LOC_Og05620 CACGAGCACCCACAATGTAG AACGGTCACCCCTTCATCTT

表2

3个氮水平下钾含量、干重和钾积累变化"

性状
Trait		 参数
Parameter		 氮水平 N level
N0 N1 N2
K含量K content (mg g-1) 均值Mean 28.87±2.85 28.05±3.35 28.05±3.20
变幅Range 22.21-35.12 20.02-37.03 19.23-38.92
干重Dry weight (g plant-1) 均值Mean 4.68±1.29 6.30±1.69 7.70±2.02
变幅Range 1.94-8.12 2.66-10.30 3.68-14.40
钾积累K accumulation (mg plant-1) 均值Mean 26.47±7.07 34.91±8.76 42.51±10.89
变幅Range 11.41-48.26 15.58-53.72 21.71-72.93

图1

3 种氮水平下水稻钾含量、干重和钾积累的分布 a、c和e分别为N0、N1和N2条件下的钾含量、干重和钾积累分布; b、d、f 为N0、N1和N2条件下钾含量、干重和钾积累分布的气泡图, 气泡宽度代表株系数, 红色虚线为3 种氮条件下各性状的平均值。"

图2

不同氮处理下钾含量和钾积累相对变化的分布 a和b是分别相对N0N1-N0和N2N2-N1钾含量和钾积累相对变化分布。KC: 钾含量; KA: 钾积累。"

图3

钾含量、干重和钾积累在3种氮水平处理下籼粳亚种间的比较 a、b、c是钾含量、干重和钾积累在3种氮水平处理下籼粳亚种间的比较。C0i、C1i、C2i分别表示籼稻在N0、N1、N2下的钾含量, C0j、C1j、C2j分别表示梗稻在N0、N1、N2下的钾含量; W0i、W1i、W2i分别表示籼稻在N0、N1、N2下的植株干重, W0j、W1j、W2j分别表示粳稻在N0、N1、N2下的植株干重; A0i、A1i、A2i分别表示籼稻在N0、N1、N2下的钾积累, A0j、A1j、A2j分别表示粳稻在N0、N1、N2下的钾积累。"

表3

参试水稻材料钾含量、干重和钾积累与氮水平的相关系数"

氮水平
N level		 钾含量
K content		 干重
Dry weight		 钾积累
K accumulation
N0 N1 N0 N1 N0 N1
N1 -0.11 0.48** 0.46**
N2 -0.14 -0.02 0.67** 0.35** 0.66** 0.37**

表4

3个氮水平下参试水稻材料3个目标性状间的相关系数"

性状
Trait		 N0 N1 N2
K含量
K content		 干重
Dry weight		 K含量
K content		 干重
Dry weight		 K含量
K content		 干重
Dry weight
干重Dry weight -0.33** -0.38** -0.30**
K积累 K accumulation 0.06 0.92** 0.09 0.88** 0.16 0.89**

表5

3种氮处理下钾含量、干重和钾积累的显著性关联位点"

性状
Trait		 氮水平
Nitrogen level		 染色体
Chr.		 物理距离
Position (bp)		 主要等位基因
Major allele		 次要等位基因
Minor allele		 最小等位基因频率
Minor allele frequency		 P-value
(-lg P)
钾积累 N0 6 21,362,789 G A 0.087 6.803
K accumulation N1 6 21,336,732 A G 0.387 6.130
钾含量 N1 1 2,908,421 A G 0.302 6.313
K content N1 2 7,907,735 A C 0.477 6.900
N1 3 5,568,543 A G 0.317 6.791
N1 6 1,524,776 G A 0.228 7.643
N1 6 26,011,593 C T 0.077 6.400
植株干重 N0 6 21,337,192 T C 0.380 6.265
Plant weight N1 6 21,337,192 T C 0.378 7.303
N1 6 23,166,060 C T 0.049 6.267
N1 10 19,108,030 C A 0.105 6.248
N2 10 19,108,030 C A 0.105 6.282

图4

3个氮水平下水稻钾含量、干重和钾积累的全基因组关联分析 a、b和c分别是3个氮水平下水稻钾含量、干重和钾积累的全基因组关联分析。"

表6

钾含量在N0和N2处理下的相对变化的显著关联位点"

性状
Trait		 染色体
Chr.		 物理距离
Position (bp)		 主要等位基因
Major allele		 次要等位基因
Minor allele		 最小等位基因频率
Minor allele frequency		 P-value
(-lg P)
KCN1-N0 9 6,934,774 T A 0.149 6.672
KCN1-N0 10 2,822,026 C T 0.066 6.296
KCN1-N0 10 4,669,497 G A 0.030 6.201
KCN2-N1 10 2,822,026 C T 0.066 6.938

图5

钾含量和钾积累在N0和N2相对变化的全基因组关联分析 a和b分别为钾含量和钾积累在N0和N2相对变化的全基因组关联分析。KC: 钾含量; KA: 钾积累。"

图6

在3个氮水平下4个候选基因相对表达水平 在3个氮水平中, 雷达图以N1水平基因的表达量为1。"

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