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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (3): 739-746.doi: 10.3724/SP.J.1006.2022.12011

• TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

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 Online:2022-03-12 Published:2021-07-19
  • Contact: ZHA Man-Rong E-mail:wy90408@163.com;zmr0729@163.com
  • 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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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

Table 1

Primers for qRT-PCR in this study"

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

Fig. 1

Screening of pOsSUT1::AtAMT1.2 transgenic lines A: the relative expression level of AtAMT1.2 in leaf; B: tiller number. *: P < 0.05; **: P < 0.01. (n = 3)."

Fig. 2

Growth of pOsSUT1::AtAMT1.2 transgenic rice plants under different nitrogen conditions in 2020 (A): phenotype of rice plants at elongation stage, Bar: 15 cm; (B) number of tillers and (C) dry weight of transgenic rice plants in HN; (D) tillers number and (E) dry weight of transgenic rice plants in LN. *: P < 0.05; **: P < 0.01 (n = 3); HN: high nitrogen; LN: low nitrogen."

Table 2

Effects of different nitrogen rates on grain yield and yield components in pOsSUT1::AtAMT1.2 transgenic rice plants"

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

Table 3

Effects of nitrogen amount on nitrogen concentration in the different organs of pOsSUT1::AtAMT1.2 transgenic rice"

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

Fig. 3

Nitrogen use efficiency (NUE) of pOsSUT1::AtAMT1.2 transgenic rice plants under different nitrogen levels The transgenic rice plants were planted in September in 2020. *: P < 0.05; **: P < 0.01 (n = 3); HN: high nitrogen; LN: low nitrogen."

Fig. 4

Tiller number and biomass accumulation of pOsSUT1::AtAMT1.2 transgenic rice plants The transgenic rice plants were planted in September in 2020. (A) the number of tillers; (B) fresh weight; (C) the relative expression levels of genes related to the strigolactone pathway in tillering buds. *: P < 0.05; **: P < 0.01 (n = 3); DAS: days after seeding."

Fig. 5

Relative expression patterns of genes related to nitrogen metabolism in pOsSUT1::AtAMT1.2 transgenic rice plant leaves under high nitrogen The transgenic rice plants were planted in September in 2020. *: P < 0.05; **: P < 0.01 (n = 3)."

Fig. 6

Carbon and nitrogen metabolism in pOsSUT1::AtAMT1.2 transgenic rice plant leaves under high nitrogen The transgenic rice plants were planted in September in 2020. (A) the photosynthetic efficiency; (B) the content of starch; (C) the content of soluble sugar; (D) the relative expression levels of sugar transporters. *: P < 0.05; **: P < 0.01 (n = 3); Pn: net photosynthetic rate."

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