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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (11): 1661-1672.doi: 10.3724/SP.J.1006.2018.01661

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Transcriptome Analysis of Premature Senescence Induced by Pollination-prevention in Maize

Lian-Cheng WU,Pei LI,Lei TIAN,Shun-Xi WANG,Ming-Na LI,Yu-Yu WANG,Sai WANG,Yan-Hui CHEN()   

  1. College of Agronomy, Henan Agricultural University / Collaborative Innovation Center of Henan Grain Crops / National Key Laboratory of Wheat and Maize Crop Science, Zhengzhou 450046, Henan, China
  • Received:2018-01-02 Accepted:2018-07-20 Online:2018-11-12 Published:2018-07-30
  • Contact: Yan-Hui CHEN E-mail:chy9890@163.com
  • Supported by:
    This study was supported by the National Key Research and Development Program of China(2016YFD0101205-3);the Basic Frontier Project of Henan Science and Technology Department(142300413218)

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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

Table 1

Primers used in RT-qPCR analysis"

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

Fig. 1

Phenotypic changes of Yu 816 under POL and Non-POL conditionsDAS: days after silking; POL: pollination; Non-POL: non-pollination."

Fig. 2

Chlorophyll content changes of ear leaves in non-pollination(Non-POL) and pollination (POL) plants** indicates extremely significant difference at P<0.01. DAS: days after silking."

Table 2

Overview of the sequencing reads"

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

Fig. 3

Principal component analysis (PCA) for all samples including three replicates N: non-pollination treatment; Y: pollination treatment; r: biological replicates."

Fig. 4

Validation of DEGs by RT-qPCR"

Fig. 5

Number of up- and down-regulated differentially expressed genes (DEGs)“10N-10Y” represents pollination treatment group at 10 DAS vs non-pollination treatment group at 10 DAS; “24N-24Y” represents pollination treatment group at 24 DAS vs non-pollination treatment group at 24 DAS; “10Y-24Y” means pollination treatment group at 24 DAS vs pollination treatment group at 10 DAS; “10N-24N” means non-pollination treatment group at 24 DAS vs non-pollination treatment group at 10 DAS."

Fig. 6

Venn diagram of DEGs identified in POL and Non-POL plants The number in non-overlapping section indicates that of special DEGs in one pairwise comparison; the number in overlapping section of DEGs means commonly expressed in different pairwise comparisons."

Fig. 7

GO analysis of differentially expressed genes"

Fig. 8

Pathway enriched analysis of DEGs"

Table 3

Premature senescence related genes induced by pollination-prevention in maize at 10 days after silking (DAS)"

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

Table 4

Carbohydrate metabolism genes in Yu 816 ear leaves at 24 days after silking (DAS)"

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)
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