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作物杂志,2023, 第1期: 38–45 doi: 10.16035/j.issn.1001-7283.2023.01.006

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

干旱胁迫及复水对马铃薯糖代谢途径中基因表达影响的研究

刘素军1,2(), 蒙美莲3, 苏日古嘎1,2()   

  1. 1内蒙古师范大学生命科学与技术学院，010022，内蒙古呼和浩特
    2内蒙古自治区高等学校生物多样性保护与可持续利用重点实验室，010022，内蒙古呼和浩特
    3内蒙古农业大学农学院，010020，内蒙古呼和浩特
  • 收稿日期:2021-09-02 修回日期:2021-12-15 出版日期:2023-02-15 发布日期:2023-02-22
  • 通讯作者: 苏日古嘎，主要从事植物生理、生态学研究，E-mail：suriguga@imnu.edu.cn
  • 作者简介:刘素军，研究方向为马铃薯栽培生理，E-mail：20170008@imnu.edu.cn
  • 基金资助:
    国家自然科学基金(31960390);内蒙古师范大学引进高层次人才科研启动项目(2017YJRC025)

Research on the Effects of Gene Expression in Sugar Metabolism Pathway of Potato by Drought Stress and Rehydration

Liu Sujun1,2(), Meng Meilian3, Suriguga 1,2()   

  1. 1College of Life Science and Technology, Inner Mongolia Normal University, Hohhot 010022, Inner Mongolia, China
    2Key Laboratory of Biodiversity Conservation and Sustainable Utilization for College and University of Inner Mongolia Autonomous Region, Hohhot 010022, Inner Mongolia, China
    3College of Agronomy, Inner Mongolia Agricultural University, Hohhot 010020, Inner Mongolia, China
  • Received:2021-09-02 Revised:2021-12-15 Online:2023-02-15 Published:2023-02-22

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摘要：

为了明确干旱胁迫对马铃薯的损伤以及复水对其保护效果，利用高通量测序技术进行了马铃薯代谢途径中基因表达研究。试验采用盆栽法，设干旱处理（40%土壤相对含水量，DT）和干旱对照（保持土壤相对含水量为70%并持续14d，CK）、复水处理（土壤相对含水量40%维持14d后恢复至土壤相对含水量的70%，RT）和复水对照（土壤相对含水量保持在70%并维持21d，RCK），研究干旱胁迫与复水处理对马铃薯体内淀粉及糖代谢途径的影响。DT与CK处理相比，有655个基因表达发生显著变化，其中与淀粉及糖代谢途径相关的有13个基因，上调表达的有9个，下调表达的有4个，涉及的酶有11个；RT与RCK处理相比，差异表达基因有644个，与淀粉及糖代谢途径相关的有5个，上调表达的有3个，下调表达的有2个，涉及的酶有4个。干旱胁迫后，淀粉及糖代谢途径中13个基因表达发生变化，涉及代谢途径中11个酶，功能与糖基水解酶、蔗糖合酶、糖基转移酶、淀粉合酶和果胶酯酶相关。复水后，有5个基因表达仍呈显著差异，影响4个酶，发生差异表达的基因数和涉及酶的数量分别减少了8和7个，下降幅度较为明显。虽然不是完全修复，但整体修复效果较好，补偿效应显著。

关键词: 马铃薯, 基因表达, 高通量测序, 淀粉, 糖代谢

Abstract:

High-throughput sequencing technique was used to study gene expression in potato metabolic pathway to clarify the damage of potato under drought stress and the protective effects of rehydration on potato. The experiment was carried out by pot method, in which drought treatment (40% soil relative water content, DT) and drought control (keep soil relative water content 70% for 14 days, CK) were set up, and in which rehydration treatment (the relative soil water content was 40% after being maintained for 14 days and then recovered to 70% of soil relative water content, RT) and rehydration control (relative soil water content was maintained at 70% for 21 days, RCK) were set up, and the effects of drought stress and rehydration treatment on starch and sugar metabolism pathways of potato were studied. The DT and CK treatments showed significant changes in the expression of 655 genes, of which 13 genes related to starch and sucrose metabolism pathways, including nine up-regulated genes and four down-regulated genes, and there were 11 enzymes involved. There were 644 differentially expressed genes between RT and RCK treatments, of which five genes related to starch and sucrose metabolism pathways, including three up-regulated genes and two down-regulated genes, and there were four enzymes involved. Changes in the expression of 13 genes in starch and sucrose metabolic pathways were occurred after drought stress, involving 11 enzymes in metabolic pathways, their functions were related to glycosyl hydrolase, sucrose synthase, glycosyl transferase, starch synthase and pectin esterase. After rehydration, the expression of five genes was still significantly different, affecting four enzymes. The number of genes with differential expression was reduced by eight, and the number of enzymes involved was reduced by seven, the decline was more obvious. Although it was not completely repaired, the overall repair effects was better and the compensation effect was significant.

Key words: Potato, Gene expression, High-throughput sequencing, Starch, Sugar metabolism

图1

不同处理间RNA-Seq相关性检查

表1

检测数据情况汇总

处理
Treatment		 原始数据
Raw read		 处理后的数据
Clean read		 Q30
(%)		 匹配数（匹配率）
Total mapped
(rate of the mapped)		 交叉匹配数（交叉匹配率）
Multiply mapped
(rate of the multiply mapped)		 精准匹配数（精准匹配率）
Uniquely mapped
(rate of the uniquely mapped)
DT 52 481 881 49 863 309 92.03 31 377 909（62.93%） 1 101 135（2.21%） 30 276 774（60.72%）
CK 51 325 932 49 651 786 94.38 32 523 133（65.50%） 1 083 605（2.18%） 31 439 528（63.32%）
RT 45 604 570 44 071 743 94.25 28 817 595（65.39%） 912 019（2.07%） 27 905 575（63.32%）
RCK 47 472 519 45 593 043 94.17 29 233 720（64.12%） 977 935（2.14%） 28 255 785（61.97%）

表2

不同处理组间发生差异表达的基因数量统计

项目Item DT vs CK RT vs RCK
差异表达基因总数Total number of differentially expressed genes 655 644
上调基因表达数量Number of up-regulated genes 349 149
下调基因表达数量Number of down-regulated genes 306 495

图2

DT对比CK和RT对比RCK维恩图

表3

淀粉及糖代谢途径中表达差异显著的基因统计

表达发生变化的基因
Genes changed expression		 对应的酶编码
Correspond
enzyme code		 上调还是下调
Up-regulated or
down-regulated		 P值
P-value		 log2FoldChange 功能描述
Functional description
PGSC0003DMG402028252 3.2.1.26 下调 0.020165 -0.81723 糖基水解酶，C、N-末端
PGSC0003DMG400013546 2.4.1.13 上调 1.96E-06 0.78233 蔗糖合酶||糖基转移酶
PGSC0003DMG400006319 3.2.1.21 上调 0.044167 0.93775 糖苷水解酶，催化结构域，活性位点
PGSC0003DMG401017546 2.4.1.15、3.1.3.12 上调 0.021277 0.57240 海藻糖磷酸酶||糖基转移酶
PGSC0003DMG400009026
 2.7.7.27
 上调
 0.041903
 0.41162
 ADP-葡萄糖焦磷酸化酶||核苷酸-二磷酸-糖转移
酶||核苷酸基转移酶||葡萄糖-1-磷酸腺苷基转移酶
PGSC0003DMG402013540 2.4.1.21 上调 8.06E-06 1.75040 淀粉合酶，催化结构域
Novel02720 2.4.1.18 上调 0.000945 1.37100 1,4-α-葡聚糖支化酶2-2，叶绿素|淀粉分支酶II
PGSC0003DMG400007782 2.4.1.1 上调 0.014013 0.51617 糖基转移酶，家族35
PGSC0003DMG400033858 2.4.1.1 上调 0.028031 0.91992
Novel02357 5.4.2.2 上调 0.000539 0.71058 磷酸葡萄糖突变酶
PGSC0003DMG400009178 3.1.1.11 下调 0.040198 -0.85467 果胶酯酶抑制剂
PGSC0003DMG400030191 3.1.1.11 下调 0.002582 -1.41130 果胶裂解酶倍数||果胶酯酶，催化
PGSC0003DMG400002085 3.1.1.11 下调 0.005801 -1.46070 果胶酯酶抑制剂||果胶裂解酶倍数

图3

干旱胁迫对淀粉及糖代谢途径的影响 红色的表示基因表达呈显著上调，绿色的表示基因表达呈显著下调。SSM：淀粉和糖代谢途径；ASNSM：氨基糖和核苷酸糖代谢；UDGA：尿苷二磷酸半乳糖；PEC：果胶；PET：果胶盐；DGT：半乳糖醛酸；ASM：抗坏血酸代谢；DGN：D-葡萄糖醛酸；βDGS：β-D-葡萄糖苷；REM：视黄醇代谢；UDGL：尿苷二磷酸-D-葡萄糖醛酸；UDX：尿苷二磷酸-D-木糖；βDXL：1,4-β-D-木聚糖；DXY：D-木糖；PGI：戊糖与葡萄糖醛酸转化；SUCE：胞外蔗糖；αDG6P：6-磷酸-α-D-葡萄糖；βDFSN：2,6-β-D-果糖基；βDFSM：26-β-D-果糖基；βDFR：β-D-果糖；KET：3-酮蔗糖；SUC：蔗糖；SUP：蔗糖；DFR:D-果糖；DGL：D-葡萄糖；βGLU：β-1,3葡聚糖；αDGL：α-D-葡萄糖；βDGC：1,4-β-D葡聚糖；CEL：纤维素；GLU：葡萄糖苷；βDG：β-D-葡萄糖；GGL：GDP-葡萄糖；CGL：CDP-葡萄糖；αDGLP：α-D-1,6-二磷酸葡萄糖；βDFP：6-磷酸-β-D-果糖；αDGP：1-磷酸α-D-葡萄糖；ADPG：腺苷二磷酸葡萄糖；DFT：D-果糖；UGL：尿苷二磷酸葡萄糖；GLA：糖原与直链淀粉；GLY：糖酵解；GLN：糖异生；αDG6P：6-磷酸-α-D-葡萄糖；THL：海藻糖；βDG6P：6-磷酸-β-D-葡萄糖；βDGP：β-D-葡萄糖-1磷酸；DG6P：6-磷酸-D-葡萄糖；TRP：6-磷酸海藻糖；TRE：海藻糖生物合成途径集合；THLE：α、α-海藻糖（细胞外）；MAT：麦芽糖；MAP：6-磷酸麦芽糖；CYR：环麦芽糖糊精；MALE：麦芽糖（细胞外）；MAL：麦芽糖糊精；STG：淀粉与糖原；DEX：糊精；αMAP：1-磷酸麦芽糖；ISO：异麦芽糖；CEB：纤维二糖；CELL：纤维三糖；CER：纤维素四糖；CEP：纤维素五糖；CEX：纤维素己糖；CET：纤维素庚糖。下同

表4

复水后呈基因表达显著差异统计

表达发生变化的基因
Genes changed expression		 对应酶编码
Correspond
enzyme code		 上调还是下调
Up-regulated or
down-regulated		 P值
P-value		 log2FoldChange 功能描述
Functional description
PGSC0003DMG402028252 3.2.1.26 下调 0.020165 -0.81723 糖基水解酶，C、N-末端
PGSC0003DMG400026428 2.4.1.14 上调 0.007217 0.82062 蔗糖磷酸合酶||糖基转移酶
PGSC0003DMG400009177 3.1.1.11 下调 0.002437 -0.70394 果胶裂解酶||果胶酯酶抑制剂
Novel02278 2.4.1.18 上调 0.021939 0.92412 1,4-α-葡聚糖支化酶3，叶绿体/淀粉体
PGSC0003DMG400022307 2.4.1.18 上调 0.010419 0.88644 α-淀粉酶||糖苷水解酶||糖基水解酶

图4

复水对淀粉及糖代谢途径的影响

图5

10个相对表达差异基因的柱状图

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