阻断授粉诱导玉米叶片提前衰老的转录组分析

doi:10.3724/SP.J.1006.2018.01661

摘要/Abstract

摘要：

衰老发生在玉米生长发育的最后阶段, 叶片适时启动衰老对玉米最终产量的形成具有至关重要的作用。本研究以玉米自交系豫816为试材, 采用RNA-seq技术分析阻断授粉诱导玉米叶片提前衰老的分子机制。与正常授粉植株相比, 阻断授粉诱导条件下的植株在吐丝后27 d, 叶片全部变黄并枯萎。吐丝后24 d, 授粉植株与同期非授粉植株叶片间的叶绿素含量差异达到极显著水平。差异表达基因(DEG)分析结果显示, 吐丝后10 d的授粉处理组与同期非授粉处理组比较存在173个DEG; 吐丝后24 d的授粉处理组与同期非授粉处理组比较存在835个DEG。吐丝后24 d的授粉处理组与吐丝后10 d的授粉处理组比较存在1381个DEG; 吐丝后24 d的非授粉处理组与吐丝后10 d的非授粉处理组比较存在1591个DEG。GO功能富集分析发现, 吐丝后10 d, 授粉处理组和非授粉处理组间DEG的功能主要富集在刺激响应和代谢进程; 吐丝后24 d, 授粉处理与非授粉处理间DEG的功能主要富集在光合作用进程。Pathway富集分析结果显示, 吐丝后10 d, 授粉处理与非授粉处理间的DEG主要参与RNA降解、光合作用、木质素合成、转录调控、糖转运代谢路径; 吐丝后24 d, 授粉处理与非授粉处理间的DEG主要参与信号传导、激素代谢和光合作用路径。阻断授粉诱导处理使豫816植株体内碳水化合物代谢和光合作用受到影响, 造成玉米衰老反应的启动和衰老速率显著提前和加快。

关键词: 玉米, 阻断授粉, 早衰, 转录组

Abstract:

Senescence occurs in the last stage of maize growth and development. Timely started leaf aging has a crucial role on the formation of maize final yield. Maize inbred line Yu 816 was used to explore the molecular mechanism of early senescence induced under pollination-prevention by transcriptome analysis. Compared with the normal pollination plants, the leaves of non-pollination plants turned yellow and withered at 27 days after silking (DAS). Leaf chlorophyll content difference between pollination and non-pollination plants reached extremely significant level at 24 DAS. RNA-seq assay revealed there were 173 and 835 differentially expressed genes (DEGs) between pollinated and non-pollinated treatments at 10 DAS and 24 DAS, respectively. There were 1381 DEGs in pollination treatment group and 1591 DEGs in non-pollination treatment group between 10 DAS and 24 DAS. GO analysis showed that DEG functions between pollination and non-pollination treatments were mostly enriched in stimulus response and metabolic process at 10 DAS, whereas mainly in photosynthesis process at 24 DAS. Furthermore, pathway enrichment analysis showed that DEGs between pollinated and non-pollinated treatments were mainly involved in the metabolic pathways such as RNA degradation, photosynthesis, lignin synthesis, transcription regulation and sugar transport at 10 DAS, while primarily in the processes of signaling, hormone metabolism, photosynthesis at 24 DAS. Carbohydrate metabolism and photosynthesis processes affected by pollination-prevention result in the senescence onset and the significantly fast aging ahead of schedule in Yu 816 plants.

Key words: maize, pollination-prevention, premature senescence, transcriptome

吴连成,李沛,田磊,王顺喜,李明娜,王宇宇,王赛,陈彦惠. 阻断授粉诱导玉米叶片提前衰老的转录组分析[J]. 作物学报, 2018, 44(11): 1661-1672.

Lian-Cheng WU,Pei LI,Lei TIAN,Shun-Xi WANG,Ming-Na LI,Yu-Yu WANG,Sai WANG,Yan-Hui CHEN. Transcriptome Analysis of Premature Senescence Induced by Pollination-prevention in Maize[J]. Acta Agronomica Sinica, 2018, 44(11): 1661-1672.

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链接本文: https://zwxb.chinacrops.org/CN/10.3724/SP.J.1006.2018.01661

https://zwxb.chinacrops.org/CN/Y2018/V44/I11/1661

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基因编号 Gene ID	正向引物 Forward primer (5°-3°)	反向引物 Reverse primer (5°-3°)
GRMZM2G033493	AATGCAACGGAGCCAACAAT	TTTGTGACAGCTTCGTTCGG
GRMZM2G035243	CCAGCCATCCGTCTATCCAT	TCTAATCTTGCAGCGCGAAC
GRMZM2G109070	GTGTACTACGAGAGGTCCGG	AAAGCCCCAAAACGCATCTT
GRMZM2G117198	GGACACATGTTCGGGTATGC	ATTGGTCACTGTCTCGTCGT
GRMZM2G339563	ATCGTTCTTCAAGGCCAGGA	CATCTCGCGCTTTGAAAGGA
GRMZM5G801627	TGTGCAGGCGACCATGTATA	CATCAACTCAAGACGCCGTT
GRMZM2G088053	AGAGTGAGGCCCAAGATGAC	CTCAGCCTCTCCATCCTCAG
GRMZM2G109627	CGAGGATAACTGCAACGGTG	GTCGTGCAGCTGATGAGAAG
GRMZM2G062129	AGCAAGTCTGATGGCTCACT	AGCCAACCCTTGACTAGCAT
GRMZM2G064962	CGAAACACCACGATCCAAGG	ATGTAGACTGCCTCCCACAC

样本 Sample	原始数据 Raw data count	过滤后的 reads数量 Count after filter	过滤后 reads占比 Reads keep rate (%)	比对上的reads数量Mapped reads	比对上的 reads占比 Mapped reads rate (%)	在参考序列上有唯一比对位置的 reads数量 Unique mapped reads	有唯一比对位置的reads占比 Unique mapped reads (%)
10DASN_r1	7442024	7291875	97.98	6731287	92.31	5480898	81.42
10DASN_r2	9376032	9181843	97.93	8479906	92.36	7106564	83.80
10DASY_r1	7941949	7777455	97.93	7134198	91.73	5833995	81.78
10DASY_r2	8834814	8644115	97.84	7960889	92.10	6607957	83.01
24DASN_r1	9382351	9163732	97.67	8367413	91.31	7079381	84.61
24DASN_r2	8201533	8054678	98.21	7433771	92.29	6190232	83.27
24DASY_r1	8599542	8432296	98.06	7728019	91.65	6398312	82.79
24DASY_r2	9545411	9339804	97.85	8557539	91.62	7181523	83.92
24DASY_r3	8206919	8027609	97.82	7381500	91.95	6095486	82.58

玉米基因编号 Maize gene ID	差异倍数 FC (10N/10Y)	基因功能 Gene function
GRMZM2G127846	9.51	Exonuclease family protein
GRMZM2G009223	6.56	Glucose-6-phosphate/phosphate translocator 2 (GPT2)
GRMZM2G339562	6.17	Response to low sulfur 4 (LSU4)
GRMZM2G087254	5.78	APS reductase 3 (APR3)
GRMZM2G071630	3.90	Glyceraldehyde-3-phosphate dehydrogenase C2
GRMZM2G345700	3.83	Bifunctional inhibitor/lipid-transfer protein/seed storage 2S albumin superfamily protein
GRMZM2G139874	3.66	Cinnamate-4-hydroxylase (C4H)
GRMZM2G000264	3.33	H(+)-ATPase 11
GRMZM2G079613	3.23	Tetratricopeptide repeat-like superfamily protein
GRMZM2G144346	0.35	B-box type zinc finger protein with CCT domain
GRMZM2G123896	0.33	Dormancy/auxin associated family protein
GRMZM2G051151	0.31	Oxidative stress 3 (OXS3)
GRMZM2G106792	0.30	NDR1/HIN1-like 2
GRMZM5G801949	0.28	Sugar transporter 4 (STP4)
玉米基因编号 Maize gene ID	差异倍数 FC (10N/10Y)	基因功能 Gene function
GRMZM2G068510	0.24	S-adenosyl-L-methionine-dependent methyltransferases superfamily protein
GRMZM2G172214	0.22	CBS domain containing membrane protein
AC208201.3_FG002	0.20	Protein phosphatase 2C family protein
GRMZM2G412601	0.19	SOS3-interacting protein 3 (SIP3)
GRMZM2G088819	0.16	Calcium-binding EF-hand family protein
GRMZM2G084958	0.15	Protochlorophyllide oxidoreductase A (PORA)
GRMZM2G478553	0.15	RING/U-box superfamily protein (ATL3)
GRMZM2G142802	0.14	Aluminium induced protein
GRMZM2G149024	0.12	Galacturonosyltransferase-like 2
GRMZM2G177050	0.11	SOS3-interacting protein 4 (SIP4)
GRMZM2G131055	0.08	Glycosyltransferase family 61 protein

玉米基因编号 Maize gene ID	差异倍数 FC (24N/24Y)	基因功能 Gene function
GRMZM2G001304	0.19	Trehalose-6-phosphate synthase (TPS)
GRMZM2G068943	0.22	Trehalose-6-phosphate synthase (TPS)
GRMZM2G112830	4.55	Trehalose-phosphatase (TPP)
GRMZM2G099860	4.04	Trehalose-phosphatase (TPP)
GRMZM2G140614	2.75	Glucose-6-phosphate isomerase
GRMZM2G076075	6.43	Glucose-6-phosphate isomerase
GRMZM2G106213	40.55	ADP glucose pyrophosphorylase (AGPase)
GRMZM2G348551	6.92	Starch synthase 2 (SS2)
GRMZM2G089136	0.07	Phosphoglycerate kinase
GRMZM2G104632	2.86	Glyceraldehyde 3-phosphate dehydrogenase (GAP-DH)
GRMZM2G345493	0.09	Fructose-bisphosphate aldolase
GRMZM2G155253	0.05	Fructose-bisphosphate aldolase
玉米基因编号 Maize gene ID	差异倍数 FC (24N/24Y)	基因功能 Gene function
GRMZM2G089365	8.92	Fructose-bisphosphate aldolase
GRMZM2G046284	0.16	Fructose-bisphosphate aldolase
GRMZM5G836250	0.11	Fructose-1,6-bisphosphatase
GRMZM2G306732	0.10	Fructose-1,6-bisphosphatase
GRMZM5G875238	0.16	Sucrose phosphate synthase
GRMZM2G466780	0.20	Fructokinase-like 1 (FLN1)
GRMZM2G103843	0.28	Fructokinase-like 2 (FLN2)