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作物学报 ›› 2022, Vol. 48 ›› Issue (3): 739-746.doi: 10.3724/SP.J.1006.2022.12011

• 耕作栽培·生理生化 • 上一篇    下一篇

增强叶片氮素输出对水稻分蘖和碳代谢的影响

王琰1(), 陈志雄2, 姜大刚3, 张灿奎4, 查满荣1,*()   

  1. 1吉首大学生物资源与环境科学学院, 湖南吉首 416000
    2华南农业大学农学院, 广东广州 510642
    3华南农业大学生命科学学院, 广东广州 510642
    4普渡大学农学院, 美国印第安纳州 47907
  • 收稿日期:2021-02-10 接受日期:2021-06-16 出版日期:2022-03-12 网络出版日期:2021-07-19
  • 通讯作者: 查满荣
  • 作者简介:E-mail: wy90408@163.com
  • 基金资助:
    国家自然科学基金项目(32060432);湖南省教育厅项目(18C0578);广东省农作物种质资源保存与利用重点实验室开放课题项目(2020B121201008)

Effects of enhancing leaf nitrogen output on tiller growth and carbon metabolism in rice

WANG Yan1(), CHEN Zhi-Xiong2, JIANG Da-Gang3, ZHANG Can-Kui4, ZHA Man-Rong1,*()   

  1. 1College of Biology and Environmental Sciences, Jishou University, Jishou 416000, Hunan, China
    2College of Agriculture, South China Agricultural University, Guangzhou 510642, Guangdong, China
    3College of Life Sciences, South China Agricultural University, Guangzhou 510642, Guangdong, China
    4Department of Agronomy, Purdue University, Indiana 47907, IN, USA
  • Received:2021-02-10 Accepted:2021-06-16 Published:2022-03-12 Published online:2021-07-19
  • Contact: ZHA Man-Rong
  • Supported by:
    National Natural Science Foundation of China(32060432);Research Foundation of Education Bureau of Hunan Province(18C0578);Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization(2020B121201008)

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摘要：

增施氮肥是保证水稻高产的重要栽培措施, 但高氮肥投入所增加的植株氮素积累大部分滞留在营养器官中, 对产量的促进作用有限。叶片是氮素储存的主要器官及籽粒氮素的主要供给源。为了明确植株中氮素分配对水稻生长的影响, 本研究将拟南芥铵转运蛋白基因AtAMT1.2在水稻韧皮部特异表达, 促进叶片氮素输出, 检测转基因水稻植株在不同氮肥浓度下的生长情况。试验结果显示, 高氮下pOsSUT1:: AtAMT1.2转基因水稻分蘖数、氮素利用效率显著增加, 叶片中糖输出量增加, 分蘖芽中独角金内酯途径相关基因OsTB1、OsD14表达水平下调。研究说明增加叶片氮素输出能够增大叶片中糖向分蘖芽的转运量, 促进分蘖生长, 从而提高了有效分蘖数并带来了更高的氮素利用效率。

关键词: 氮素, 水稻, 分蘖, 氮素利用效率

Abstract:

Nitrogen fertilizer application is one of the main cultivation measures to raise the yield, and high nitrogen level has limited contribution to grain yield due to limited nitrogen translocation in rice. To clarify the effects of nitrogen allocation on rice growth, we constructed pOsSUT1::AtAMT1.2 transgenic rice, the ammonium transporter gene AtAMT1.2 specific expression in phloem to promote leaf nitrogen output. The growth and yield of transgenic plants were measured under HN (high nitrogen) and LN (low nitrogen) conditions. Compared to WT plants, more tillers and higher grain yield were detected in transgenic plants in response to HN condition. The sugar output in leaves was increased, and the relative expression levels of the strigolactone pathway related genes OsTB1 and OsD14 in tiller buds were down-regulated. Our results indicated that the increase of leaf nitrogen export by overexpressing AtAMT1.2 gene could promote sugar translocation from leaves to tillering buds, which improved the growth of tiller, increased the effective tiller number and nitrogen use efficiency.

Key words: nitrogen, rice (Oryza sativa L.), tiller, nitrogen use efficiency

表1

荧光定量PCR引物序列"

基因名称
Gene name		 正向引物
Forward primer (5'-3')		 反向引物
Reverse primer (5'-3')
OsSUT1 TCATCCCTCAGGTGGTCATCG CTTGGAGATCTTGGGCAGCAG
OsSUT2 GTCATACCACAGGTTATTGTGTC GAATTGCAAAGAATGGCCG
OsSUT4 CGTTGTTCCGCAGATAGTAGTG GTGTTCTGCTCAGCCAAATCC
OsSSI GGGCCTTCATGGATCAACC CCGCTTCAAGCATCCTCATC
OsGBSSI AACGTGGCTGCTCCTTGAA TTGGCAATAAGCCACACACA
OsGBSSIl AGGCATCGAGGGTGAGGAG CCATCTGGCCCACATCTCTA
OsFd-GOGAT TGGTTGAGGGCACTGGAGATCA AATATAGGCAAGGCCACCCGTC
OsNADH-GOGAT CCTGTCGAAGGATGATGAAGGTGA TGCATGGCCCTACTATCTTCGC
OsGS1.1 CAAGTCTTTTGGGCGTGATAT CTCAAGAATGTAGCGAG
OsGDH1 CATCTGATCATCTCCCTGTT TTCAGGCAATTCATCACTAC
OsGDH2 GGCCATTAACAACACTCATA ACGCCGATCTATCTTGAAT
OsGDH3 CCAAAAGTACATGAAGAACG GTGATTCCTCAACAGATTCTC
OsTB1 GCCGGATGCAAGAAATC TCAGCAGTAGTGCCGCGAA
OsD14 CGCCTTCGTCGGCCACTC TCGAACCCGCCGTGGTAGTC
OsMADS57 ATGGGGAGGGGGAAGATAG AATTTAGGCTTCTAGAAAGTTCG
AtAMT1.2 ATGGCGACGTGCTTGGACAG CGAGCACGTTGGTGAGCATG
Actin CAATCGTGAGAAGATGACCC GTCCATCAGGAAGCTCGTAGC

图1

pOsSUT1::AtAMT1.2转基因水稻株系筛选 A: AtAMT1.2在叶片中表达量; B: 分蘖数。*和**分别表示在0.05和0.01水平上显著(n = 3)。"

图2

pOsSUT1::AtAMT1.2转基因水稻植株在不同氮素条件下的生长情况 拔节期水稻表型(A), 标尺为15 cm; 高氮下分蘖数(B)、干重(C); 低氮下分蘖数(D)、干重(E)。*和**分别表示在0.05和0.01水平上显著(n = 15); HN: 高氮; LN: 低氮。"

表2

施氮量对pOsSUT1::AtAMT1.2转基因水稻产量及其构成因素的影响"

种植时间/处理
Planting time/treatment		 品种
Cultivar		 有效穗数/株
Effective panicle per plant		 颖花数/穗
Spikelets per panicle		 颖花数/株
Spikelets per plant		 产量/株
Grain yield (g plant-1)
2020/5 高氮HN WT 7.0±1.9 72.3 ±11.9 490.5±95.3 9.8±1.8
SA11 10.0±2.1** 74.5±11.2 721.3±140.1** 16.6±2.5**
SA33 9.6±2.2* 75.8±12.2 744.3±152.8** 17.0±2.5**
低氮LN WT 5.3±1.5 52.5±7.1 246.6±60.8 5.4±1.0
SA11 5.3±1.6 52.4±6.4 257.4±44.1 5.5±1.1
SA33 5.5±1.5 53.3±8.8 255.2±47.5 5.5±1.0
2020/9 高氮HN WT 9.6±1.0 83.7±17.2 833.1±114.0 19.6±2.3
SA11 15.2±1.4** 82.5±10.1 1146.7±214.4** 27.9±3.9**
SA33 14.1±1.5* 86.7±17.4 1087.6±146.3* 27.5±4.0**
低氮LN WT 6.5±1.0 61.7±8.0 403.8±77.8 9.7±1.7
SA11 6.9±0.9 63.1±8.9 442.6±75.1 10.0±1.4
SA33 6.7±1.1 62.1±7.9 420.4±73.6 9.8±0.9

表3

施氮量处理对pOsSUT1::AtAMT1.2转基因水稻各器官氮浓度的影响"

种植时间/处理
Planting date/treatment		 品种
Cultivar		 叶
Leaf (%)		 茎
Steam (%)		 穗
Panicle (%)
2020/5 高氮HN WT 1.07±0.09 0.96±0.06 1.03±0.08
SA11 0.98±0.05* 1.01±0.10 1.15±0.07**
SA33 0.97±0.07* 1.02±0.07* 1.13±0.08**
低氮LN WT 0.86±0.05 0.83±0.06 0.79±0.06
SA11 0.85±0.04 0.86±0.04 0.81±0.05
SA33 0.87±0.06 0.85±0.04 0.82±0.06
2020/9 高氮HN WT 1.00±0.09 0.94±0.06 1.12±0.12
SA11 0.93±0.04* 1.01±0.10 1.28±0.11**
SA33 0.91±0.08* 1.04±0.11* 1.27±0.09**
低氮LN WT 0.84±0.08 0.83±0.06 0.81±0.06
SA11 0.85±0.07 0.84±0.05 0.84±0.11
SA33 0.83±0.09 0.86±0.06 0.83±0.06

图3

pOsSUT1::AtAMT1.2转基因水稻植株在不同氮肥条件下的氮肥农学利用率 转基因水稻植株为2020年9月种植批次。*和**分别表示在0.05 和0.01 水平上显著(n = 3); HN: 高氮; LN: 低氮。"

图4

pOsSUT1::AtAMT1.2转基因水稻植株分蘖数及生物量积累动态 转基因水稻植株为2020年9月种植批次。(A)分蘖数; (B)鲜重; (C)分蘖芽中独角金内酯途径相关基因表达量。*和**分别表示在0.05 和0.01 水平上显著(n = 3); DAS: 播种后天数。"

图5

高氮下pOsSUT1::AtAMT1.2转基因水稻叶片中氮代谢相关基因表达量 转基因水稻植株为2020年9月种植批次。*和**分别表示在0.05和0.01 水平上显著(n = 3)。"

图6

高氮下pOsSUT1::AtAMT1.2转基因水稻叶片中碳氮代谢变化 转基因水稻植株为2020年9月种植批次。(A)光合效率; (B)淀粉含量; (C)可溶性糖含量; (D)糖转运相关基因表达量。*和**分别表示在0.05和0.01水平上显著(n = 3); Pn: 净光合速率。"

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