不同氮肥水平下水稻倒二叶叶鞘的转录组分析

doi:10.16819/j.1001-7216.2025.250306

中国水稻科学 ›› 2025, Vol. 39 ›› Issue (4): 563-574.DOI: 10.16819/j.1001-7216.2025.250306

• 研究报告 • 上一篇

不同氮肥水平下水稻倒二叶叶鞘的转录组分析

黄福灯¹^,^#, 吴春艳¹^,^#, 郝媛媛¹, 韩一飞¹, 张小斌¹^,^*(), 孙会锋²^,^*(), 潘刚³

¹浙江省农业科学院, 杭州 310021
²海盐县农业技术推广中心，浙江嘉兴 314300
³浙江大学农业与生物技术学院, 杭州 310058

收稿日期:2025-02-17 修回日期:2025-04-11 出版日期:2025-07-10 发布日期:2025-07-21
通讯作者: ^*email: 525588094@qq.com,zhangxb@zaas.ac.cn
作者简介:
^#共同第一作者
基金资助:
浙江省水稻新品种选育重大科技专项(2021C02063-1);浙江省农业科学院重大科技任务攻关项目(2024ZDZX01)

Transcriptome Analysis of Top Second Leaf Sheath of Rice Under Different Nitrogen Fertilizer Levels

HUANG Fudeng¹^,^#, WU Chunyan¹^,^#, HAO Yuanyuan¹, HAN Yifei¹, ZHANG Xiaobin¹^,^*(), SUN Huifeng²^,^*(), PAN Gang³

¹Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
²Haiyan Agricultural Technology Promotion Center, Jiaxing, 314300, China
³College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China

Received:2025-02-17 Revised:2025-04-11 Online:2025-07-10 Published:2025-07-21
Contact: ^*email: 525588094@qq.com,zhangxb@zaas.ac.cn
About author:
^＃These authors contributed equally to this work

摘要/Abstract

摘要：

【目的】探明叶鞘对氮素响应的重要基因，为水稻氮素高效利用的遗传改良提供科学依据。【方法】将粳稻品种秀水134种植在低氮(LN)、中氮(MN)和高氮(HN)的渗漏池中，选取抽穗后第10天的倒二叶叶鞘进行转录组测序分析，以鉴定叶鞘对氮素响应的重要基因。【结果】转录组数据比较分析显示，三种氮素水平间存在1791个差异表达基因。其中HN与MN、MN与LN、以及HN与LN组间的上调和下调表达基因数分别为312个和155个，263个和160个，1059个和542个。三种施氮水平间的共有上调和下调差异表达基因分别为15个和53个，涉及营养元素吸收和转运、生物与非生物胁迫响应、植物激素响应以及光调控等生理生化过程。基于GO富集分析和KEGG代谢途经分析，最终鉴定出参与氮素利用及光合作用的已知功能基因分别为6个和4个，以及50个功能未知基因，推测这些基因在倒二叶叶鞘的氮素响应中发挥重要作用。【结论】研究结果初步明确了叶鞘对不同氮素水平响应的重要基因，这些基因在氮素利用的信号转导途径中发挥重要作用。

关键词: 水稻, 倒二叶叶鞘, 氮肥, 转录组分析

Abstract:

【Objective】 The leaf sheath is an important organ that simultaneously serves the functions of “source, sink, and transport”, playing a crucial role in the growth and development of rice plants. Identifying important genes in leaf sheaths in response to nitrogen will provide scientific evidence for the genetic improvement of nitrogen use efficiency in rice. 【Method】 The japonica rice cultivar Xiushui 134 was planted in percolation ponds at low nitrogen (LN), medium nitrogen (MN), and high nitrogen (HN) levels. The top second leaf sheaths were collected on the 10th day after heading for transcriptome sequencing analysis to identify important genes involved in the nitrogen response of rice leaf sheaths. 【Result】 Comparative analysis of the transcriptome data revealed 1,791 differentially expressed genes at the three nitrogen levels. The number of upregulated and downregulated genes between HN and MN, MN and LN, and HN and LN were 312 and 155, 263 and 160, and 1,059 and 542, respectively. The common upregulated and downregulated genes at the three nitrogen levels were 15 and 53, respectively, involved in physiological and biochemical processes such as nutrient element absorption and transport, responses to biotic and abiotic stresses, plant hormone responses, and light regulation. Based on GO enrichment analysis, KEGG metabolic pathway analysis, a total of 6 known functional genes related to nitrogen utilization and 4 genes related to photosynthesis, as well as 50 genes of unknown function, were identified. It is speculated that these genes play important roles in the nitrogen response of the second leaf sheath in rice. 【Conclusion】 The research results preliminarily clarify the important genes involved in the leaf sheath's response to different nitrogen levels, which may play significant roles in signal transduction pathways of nitrogen utilization.

Key words: rice, top second leaf sheath, nitrogen fertilizer, transcriptome analysis

黄福灯, 吴春艳, 郝媛媛, 韩一飞, 张小斌, 孙会锋, 潘刚. 不同氮肥水平下水稻倒二叶叶鞘的转录组分析[J]. 中国水稻科学, 2025, 39(4): 563-574.

HUANG Fudeng, WU Chunyan, HAO Yuanyuan, HAN Yifei, ZHANG Xiaobin, SUN Huifeng, PAN Gang. Transcriptome Analysis of Top Second Leaf Sheath of Rice Under Different Nitrogen Fertilizer Levels[J]. Chinese Journal OF Rice Science, 2025, 39(4): 563-574.

图/表 11

参考文献 31

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基因 ID	正向引物 Forward primer(5’-3’)	反向引物 Reverse primer(5’-3’)
LOC_Os07g48020	GCTCCAAGGTGAACTCCTAATT	CACCGTACCTATACTTGTGGTT
LOC_Os07g08160	GCCCTTCGTCAACGTCTAATTA	CCCTAGCTAGCTACCCAGTATA
LOC_Os11g25260	CAGTTACTTGCATTATGGGCTC	ATGTACCATTGAATTCGCGAAG
LOC_Os09g12290	AGTGGAAAGAGCTTCTACGAAA	TCATAATAGTGGCATGCTACGT
LOC_Os07g08150	GTACTTGCGTATGTACTGGGTA	CACTATCAAGCTTGCTACTTGC
LOC_Os01g14410	CTTCAATTCGATCCTCGCTAAG	CGCCATTGCAAATAACTCAAAC
LOC_Os10g01080	AAACACAAACACTGACGCC	GAGAAGCTACCGGACTTGTG
LOC_Os02g02120	GCCATAAAGCGCTCAAAGATTA	CTTCCACTTCTAAGCAACAACC
LOC_Os04g27670	CATTTTGAGTCTGTATGCGTGA	TGCTGCTTTTCTGTTTACTTCC

基因 ID	正向引物 Forward primer(5’-3’)	反向引物 Reverse primer(5’-3’)
LOC_Os07g48020	GCTCCAAGGTGAACTCCTAATT	CACCGTACCTATACTTGTGGTT
LOC_Os07g08160	GCCCTTCGTCAACGTCTAATTA	CCCTAGCTAGCTACCCAGTATA
LOC_Os11g25260	CAGTTACTTGCATTATGGGCTC	ATGTACCATTGAATTCGCGAAG
LOC_Os09g12290	AGTGGAAAGAGCTTCTACGAAA	TCATAATAGTGGCATGCTACGT
LOC_Os07g08150	GTACTTGCGTATGTACTGGGTA	CACTATCAAGCTTGCTACTTGC
LOC_Os01g14410	CTTCAATTCGATCCTCGCTAAG	CGCCATTGCAAATAACTCAAAC
LOC_Os10g01080	AAACACAAACACTGACGCC	GAGAAGCTACCGGACTTGTG
LOC_Os02g02120	GCCATAAAGCGCTCAAAGATTA	CTTCCACTTCTAAGCAACAACC
LOC_Os04g27670	CATTTTGAGTCTGTATGCGTGA	TGCTGCTTTTCTGTTTACTTCC

性状 Trait	低氮 LN	中氮 MN	高氮 HN
株高 Plant height(cm)	78.07±0.64 b	81.67±0.88 a	83.43±1.03 a
千粒重 1000-grain weight(g)	13.69±0.08 c	21.30±0.81 a	17.90±1.12 b
单株有效穗数 Effective pancle number	8.33±0.33 a	8.67±0.33 a	9.00±0.58 a
穗干质量 Panicle dry weight(g)	3.62±0.10 a	1.86±0.42 b	1.51±0.05 b
每穗总粒数 Grain number per panicle	99.67±1.45 c	152.00±1.15 b	166.67±1.76 a
结实率 Seed-setting rate(%)	77.27±9.87 b	92.40±2.00 a	93.08±1.34 a

性状 Trait	低氮 LN	中氮 MN	高氮 HN
株高 Plant height(cm)	78.07±0.64 b	81.67±0.88 a	83.43±1.03 a
千粒重 1000-grain weight(g)	13.69±0.08 c	21.30±0.81 a	17.90±1.12 b
单株有效穗数 Effective pancle number	8.33±0.33 a	8.67±0.33 a	9.00±0.58 a
穗干质量 Panicle dry weight(g)	3.62±0.10 a	1.86±0.42 b	1.51±0.05 b
每穗总粒数 Grain number per panicle	99.67±1.45 c	152.00±1.15 b	166.67±1.76 a
结实率 Seed-setting rate(%)	77.27±9.87 b	92.40±2.00 a	93.08±1.34 a

基因 ID	基因注释 Gene annotation	FPKM
基因 ID	基因注释 Gene annotation	LN	MN	HN
LOC_Os07g08150	早期光诱导蛋白Early light-induced protein Early light-induced protein	300.51	95.23	11.46
LOC_Os01g14410	早期光诱导蛋白Early light-induced protein	372.04	143.45	24.47
LOC_Os10g01080	SOR/SNZ家族蛋白SOR/SNZ family protein	163.38	70.66	22.57
LOC_Os07g08160	早期光诱导蛋白Early light-induced protein	268.08	103.47	14.41
LOC_Os11g01210	PPR蛋白Pentatricopeptide repeat protein	23.31	14.53	5.44
LOC_Os12g01210	PPR蛋白Pentatricopeptide repeat protein	18.86	11.60	4.45
LOC_Os01g45110	花青素-3-O-β-葡萄糖苷转移酶Anthocyanin 3-O-beta-glucosyltransferase	4.93	1.72	0.75
LOC_Os01g53330	花青素-5,3-O-糖苷转移酶Anthocyanidin 5,3-O-glucosyltransferase	16.97	7.89	2.66
LOC_Os09g26580	表达蛋白Expressed protein	21.29	15.22	6.63
LOC_Os07g06800	3-酮-5-α-类固醇-4-脱氢酶3-oxo-5-alpha-steroid 4-dehydrogenase	17.75	5.66	2.04
LOC_Os03g17460	IN2-1蛋白	2.37	1.20	0.42
LOC_Os07g44110	细胞色素P450 72A1 Cytochrome P450 72A1	1.37	0.51	0.34
LOC_Os07g04820	蛋白激酶Protein kinase	1.37	0.76	0.21
LOC_Os08g07730	转移酶家族蛋白Transferase family protein	4.16	2.26	0.74
LOC_Os08g10010	酰基去饱和酶Acyl-desaturase	0.79	0.60	0.10

不同氮肥水平下水稻倒二叶叶鞘的转录组分析

Transcriptome Analysis of Top Second Leaf Sheath of Rice Under Different Nitrogen Fertilizer Levels

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Metrics

基因 ID	基因注释 Gene annotation	FPKM
基因 ID	基因注释 Gene annotation	LN	MN	HN
LOC_Os03g58290	色氨酸合成酶Indole-3-glycerol phosphate lyase	0.21	2.53	13.95
LOC_Os03g58300	色氨酸合成酶Indole-3-glycerol phosphate lyase	0.00	0.95	2.28
LOC_Os07g48020	过氧化物酶前体Peroxidase precursor	12.80	31.09	98.47
LOC_Os11g18366	环阿屯醇合酶Cycloartenol synthase	0.91	4.17	7.69
LOC_Os02g02120	类受体蛋白激酶OsWAK receptor-like protein kinase	3.88	13.72	24.3
LOC_Os02g17400	富含亮氨酸的重复蛋白Leucine rich repeat protein	0.64	1.55	4.18
LOC_Os09g12290	双功能天冬氨酸激酶Bifunctional aspartokinase/homoserine dehydrogenase	2.72	3.89	13.90
LOC_Os11g25260	核苷酸三磷酸酶Nucleoside-triphosphatase	2.65	5.47	15.06
LOC_Os06g35940	类植物开花控制同源基因FT-like12 homologous to Flowering Locus T gene	0.08	1.01	3.71
LOC_Os04g27670	萜烯合酶家族Terpene synthase family	15.00	36.16	136.51
LOC_Os05g47540	含保守肽的uORF Conserved peptide uORF-containing transcript	0.00	1.26	2.60
LOC_Os05g48200	谷氨酸合成酶Glutamate synthase	5.05	21.13	54.38
LOC_Os07g35340	DUF26激酶DUF26 kinases	0.27	1.16	2.72
LOC_Os07g45550	表达蛋白Expessed protein	0.50	1.64	3.32
LOC_Os01g03310	胰酶抑制剂前体Bran trypsin inhibitor precursor	0.78	3.10	10.34
LOC_Os05g28740	含广泛逆境蛋白结构域的蛋白Universal stress protein domain containing protein	35.66	72.48	195.92
LOC_Os11g40970	类受体蛋白激酶前体Receptor-like protein kinase precursor	1.73	4.37	11.32
LOC_Os07g46280	β-甘露糖苷酶Beta-mannosidase	0.75	4.60	11.13
LOC_Os12g43380	塔乌马提Thaumatin	0.86	3.06	9.25
LOC_Os08g30510	表达蛋白Expessed protein	8.73	17.48	76.58
LOC_Os11g25330	核苷酸三磷酸酶Nucleoside-triphosphatase	1.39	2.71	7.83
LOC_Os08g39840	脂氧酶Lipoxygenase	1.27	4.13	7.16
LOC_Os01g49320	糖基水解酶Glycosyl hydrolase	0.19	0.75	2.63
LOC_Os04g49210	2-氧草酰乙酸-3-双加氧酶2-oxoglutarate 3-dioxygenase	0.12	1.37	4.13
LOC_Os01g32364	β-甘露糖苷酶Beta-mannosidase	0.80	1.71	3.81
LOC_Os11g24140	类质体蓝素结构域蛋白Plastocyanin-like domain containing protein	5.27	10.93	66.45
LOC_Os01g66860	丝氨酸/苏氨酸蛋白激酶Serine/threonine protein kinase	0.97	5.02	16.31
LOC_Os10g02880	O-甲基转移酶O-methyltransferase	0.18	1.44	2.72
LOC_Os01g58910	生长素诱导蛋白Auxin-induced protein	0.73	2.02	4.53
LOC_Os05g47790	表达蛋白Expessed protein	0.05	0.28	0.73
LOC_Os04g52780	富亮氨酸重复受体蛋白激酶前体Leucine-rich repeat receptor protein kinase precursor	0.07	0.39	1.19
LOC_Os03g17200	植物特异结构域 TIGR01589家族蛋白Plant-specific domain TIGR01589 family protein	3.52	8.74	44.84
LOC_Os12g24020	含类硫化酶结构域的蛋白Rhodanese-like domain containing protein	0.37	0.73	3.84
LOC_Os09g08130	吲哚-3-甘油磷酸合酶Indole-3-glycerol phosphate synthase	0.46	1.02	1.98
LOC_Os05g46350	含IQ钙调素基序结构域的蛋白IQ calmodulin-binding motif domain containing protein	0.34	1.55	3.96
LOC_Os04g57350	含EH结构域蛋白EH domain-containing protein	0.26	0.92	2.02
LOC_Os06g22290	含豆科凝集素β结构域的蛋白Legume lectins beta domain containing protein	0.11	0.36	0.84

基因 ID	基因名称 Gene annotation	平均FPKM
基因 ID	基因名称 Gene annotation	低氮 LN	中氮 MN	高氮 HN
N循环相关基因Genes related to the nitrogen cycle
LOC_Os05g48200	OsNADH-GOGAT2	7.11	18.76	49.13
LOC_Os04g43070	OsAMT1;1	287.99	193.10	107.06
LOC_Os01g36720	OsNRT2.4	28.62	24.52	12.76
LOC_Os08g36480	OsNIA1	44.65	33.48	21.89
LOC_Os08g36500	OsNIA2	28.34	10.90	9.95
LOC_Os04g37500	OsGAD2	1.80	2.30	5.46
光合作用相关基因Genes related to photosynthesis
LOC_Os01g64960	psbS1	602.97	386.64	217.50
LOC_Os04g59440	psbS2	1,045.47	720.18	251.92
LOC_Os04g58200	OsPORA	6.22	9.59	25.38
LOC_Os01g14410	OsELIP	577.93	164.30	24.47

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