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作物学报 ›› 2023, Vol. 49 ›› Issue (7): 1785-1798.doi: 10.3724/SP.J.1006.2023.24137

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

转录组与代谢组联合分析揭示遮阴胁迫下甘薯的代谢响应途径

王雁楠1(), 陈金金1, 卞倩倩1, 胡琳琳2, 张莉3, 尹雨萌1, 乔守晨1, 曹郭郑1, 康志河1, 赵国瑞1, 杨国红1, 杨育峰1,*()   

  1. 1河南省农业科学院粮食作物研究所, 河南郑州 450002
    2郑州大学农学院, 河南郑州 450001
    3河南省农业科学院农业经济与信息研究所, 河南郑州 450002
  • 收稿日期:2022-06-08 接受日期:2022-11-25 出版日期:2023-07-12 网络出版日期:2022-12-28
  • 通讯作者: *杨育峰, E-mail: yyfyyf5@163.com
  • 作者简介:E-mail: alman001@qq.com
  • 基金资助:
    本研究由河南省自然科学基金项目(212300410170);河南省科技攻关项目(212102110251);河南省农业良种联合攻关项目(2022010401-2);河南省现代农业产业技术体系建设专项资金(HARS-22-04-G1);财政部和农业农村部国家现代农业产业技术体系建设专项(甘薯);财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-10-C14)

Integrated analysis of transcriptome and metabolome reveals the metabolic response pathways of sweetpotato under shade stress

WANG Yan-Nan1(), CHEN Jin-Jin1, BIAN Qian-Qian1, HU Lin-Lin2, ZHANG Li3, YIN Yu-Meng1, QIAO Shou-Chen1, CAO Guo-Zheng1, KANG Zhi-He1, ZHAO Guo-Rui1, YANG Guo-Hong1, YANG Yu-Feng1,*()   

  1. 1Cereal Crops Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
    2School of Agricultural Sciences, Zhengzhou University, Zhengzhou 450001, Henan, China
    3Institute of Agricultural Economics and Information, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan, China
  • Received:2022-06-08 Accepted:2022-11-25 Published:2023-07-12 Published online:2022-12-28
  • Contact: *E-mail: yyfyyf5@163.com
  • Supported by:
    The Natural Science Foundation of Henan Province(212300410170);The Science and Technology Research Program of Henan Province(212102110251);The Henan Joint Research Program for Improved Agricultural Varieties(2022010401-2);The Special Fund for Henan Agriculture Research System(HARS-22-04-G1);The China Agriculture Research System of MOF and MARA(甘薯);The China Agriculture Research System of MOF and MARA(CARS-10-C14)

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摘要：

甘薯是喜光作物, 但其在套种栽培模式中一般处于低位被遮阴, 大田生长中后期也时常面临阴雨寡照天气而影响块根干物质积累, 因此, 解析甘薯在遮阴胁迫下的代谢响应途径可为其耐荫性品种改良提供理论依据。本研究对甘薯品种郑红23号进行透光率50%的遮阴胁迫15 d后发现, 遮阴胁迫下郑红23号的叶绿素b以及总叶绿素含量较自然光照下均显著提高; 叶绿素光系统PSII最大光化学效率(Fv/Fm)、PSII潜在活性(Fv/Fo)和光合性能综合指数(PIABS)在遮阴胁迫下均显著下降; 净光合速率和水分利用率显著降低, SOD酶和POD酶活性则显著提高; 此外, 遮阴胁迫显著提高了郑红23号的蔓长和比叶面积, 根鲜重则显著降低。对遮阴胁迫和自然光照条件下的叶片组织进行转录组和代谢组联合分析发现, 差异基因和差异代谢物主要共同富集于苯丙素合成途径、糖代谢相关途径、鞘脂代谢途径和精氨酸合成途径。苯丙素合成途径富集到的上调差异表达基因多数为POD酶家族基因, 说明遮阴胁迫触发了甘薯的ROS活性氧清除系统。同时, 遮阴胁迫降低了甘薯植株的糖代谢水平, 叶片可溶性糖含量下降, 淀粉合成与降解均受到抑制, 块根膨大受阻。而鞘脂及精氨酸代谢途径则可能通过提高生物膜的稳定性以及增加多胺类抗逆因子的合成底物来使植株更好地适应遮阴胁迫。以上结果为理解遮阴胁迫下甘薯的代谢响应途径提供了新的理论依据。

关键词: 甘薯, 遮阴胁迫, 转录组, 代谢组, 响应途径

Abstract:

Sweetpotato is a heliophile crop. However, it is usually shaded in the lower position in the interplanting cultivation mode. During the middle and late field growing period, it often faces rainy weather with little illumination, which affects the dry matter accumulation in tuberous roots. Thus, analyzing the metabolic response pathways of sweetpotato under shade stress will provide the theoretical basis for the varieties’ genetic improvement of shade tolerance. In this study, the sweetpotato variety Zhenghong 23 was exposed to shade stress with 50% light transmittance for 15 days. Results showed that chlorophyll b and the total chlorophyll contents of Zhenghong 23 under shade stress were significantly increased compared with those under natural light. The maximum photochemical efficiency (Fv/Fm), the potential activity (Fv/Fo), and the comprehensive index of photosynthetic performance (PIABS) of the chlorophyll photosystem PSII decreased significantly under shade stress. The net photosynthetic rate and water use efficiency decreased significantly, while SOD and POD enzyme activities increased significantly. In addition, shade stress increased significantly the vine length and specific leaf area of Zhenghong 23, but reduced significantly the fresh weight of roots. Transcriptome and metabolome analysis of leaf tissues under shade stress and natural light conditions showed that the DEGs and DMs were mainly enriched in phenylpropanoid biosynthesis, sugar metabolism, sphinolipid metabolism, and arginine biosynthesis pathways. Most of the up-regulated DEGs enriched in the phenylpropanoid biosynthesis pathway were POD enzyme family genes, indicating that the shade stress triggered the ROS scavenging system in sweetpotato. Meanwhile, shade stress reduced sugar metabolism level of sweetpotato, decreased the soluble sugar content of leaves, inhibited both the synthesis and degradation of starch, and blocked the expansion of tuberous roots. In addition, the sphinolipid and arginine metabolism pathways may better adapt sweetptoato plants to shade stress through improving the stability of biomembranes and increase the synthetic substrates of polyamine anti-stress factors. These results provide new theoretical basis for understanding the metabolic response pathways of sweetpotato under shade stress.

Key words: sweetpotato, shade stress, transciptome, metabolome, the response pathways

图1

郑红23号自然光照与遮阴胁迫下的植株形态对比"

表1

遮阴胁迫对郑红23号植株形态指标及叶片酶活的影响"

光处理
Light
treatment		 蔓长
Vine length
(cm)		 茎节数
Stem internode number		 比叶面积
Specific leaf area
(cm2 g-1)		 根鲜重
Root fresh weight (g)		 SOD活性
SOD activity
(U g-1 FW)		 POD活性
POD activity
(U g-1 FW)
光照Light 43.5 (6.9) 13.3 (0.5) 211.9 (28.3) 17.9 (9.9) 339.7 (80.2) 1096.1 (230.0)
遮阴Shade 65.7 (9.3)** 14.8 (2.1) 457.6 (39.2)*** 1.2 (0.5)* 547.2 (72.8)** 2366.9 (398.0)**

表2

遮阴胁迫对郑红23号叶绿素含量(mg g-1 FW)及荧光特性的影响"

光处理
Light treatment		 叶绿素a
Chlorophyll a		 叶绿素b
Chlorophyll b		 叶绿素a/b
Chlorophyll a/b		 总叶绿素
Total chlorophyll		 Fv/Fm Fv/Fo PIABS
光照Light 1.08 (0.11) 0.45 (0.12) 2.44 (0.34) 1.53 (0.23) 0.749 (0.022) 2.838 (0.210) 0.570 (0.055)
遮阴Shade 1.21 (0.04) 0.73 (0.05)** 1.66 (0.15)** 1.94 (0.05)* 0.670 (0.033)** 2.107 (0.364)* 0.398 (0.046)**

表3

遮阴胁迫对郑红23号光合参数的影响"

光处理
Light
treatment		 净光合速率
Net photosynthetic rate
(μmol m-2 s-1)		 蒸腾速率
Transpiration
rate
(mmol m-2 s-1)		 胞间CO2浓度
Intercellular CO2 concentration
(μmol mol-1)		 气孔导度
Stomatal
conductance
(mmol m-2 s-1)		 水汽压亏缺
Vapor pressure deficit
(mb)		 水分利用效率
Water use
efficiency
(%)
光照Light 9.13 (0.87) 2.29 (0.53) 241.8 (28.2) 103.5 (8.9) 2.07 (0.18) 4.07 (0.54)
遮阴Shade 2.45 (0.54)*** 1.93 (0.46) 341.0 (13.7)** 97.3 (10.1) 2.18 (0.23) 1.02 (0.37)***

图2

RNA-seq样品相关性热图(a)及代谢组各样品间的主成分分析(b) S: 遮阴; L: 光照。"

表4

样品RNA-seq质量"

样品
Sample name		 原始序列数
Raw
reads		 过滤后序列及占比
Clean reads and
percentage (%)		 比对到基因组上的reads数及占比
Number and proportion of reads
on the genome (%)		 单一位置reads数及百分比
Unique mapped reads
and percentage (%)		 Q30比例
Q30 ratio
(%)
S1 46,514,384 45,395,970 (97.60) 32,813,096 (72.28%) 31,148,789 (68.62%) 92.72
S2 45,595,318 44,524,818 (97.65) 32,073,379 (72.03%) 30,310,383 (68.08%) 93.15
S3 48,243,946 46,619,848 (96.63) 33,903,203 (72.72%) 32,580,570 (69.89%) 93.16
S4 45,242,868 43,886,694 (97.00) 31,090,577 (70.84%) 29,766,110 (67.82%) 92.72
L1 51,826,126 50,453,206 (97.35) 36,601,089 (72.54%) 35,349,491 (70.06%) 92.79
L2 52,046,026 50,516,688 (97.06) 35,620,398 (70.51%) 34,152,030 (67.61%) 93.27
L3 43,082,564 42,246,862 (98.06) 29,421,358 (69.64%) 28,373,138 (67.16%) 92.76
L4 45,214,902 43,699,856 (96.65) 30,949,611 (70.82%) 29,612,170 (67.76%) 93.30

图3

遮阴胁迫和自然光照下鉴定出的表达基因数(a)和差异表达基因数(b) S: 遮阴; L: 光照。"

图4

差异表达基因GO富集中显著性排名前30的功能类(a)和KEGG富集中显著性排名前20的通路(b) BP: 生物过程; CC: 细胞组成; MF: 分子功能。柱上数字代表富集到的差异基因个数。纵坐标为显著性水平, 数值越高越显著。"

图5

Top20差异代谢物相关性图(a)及差异代谢物KEGG富集气泡图(b) (a) 红色代表正相关, 蓝色代表负相关, 没有颜色的点表示无显著相关性(P > 0.05)。(b) 气泡颜色与大小代表富集可信度及富集到的差异代谢物数目。-log10(P-value)越大, 富集可信度越高。S: 遮阴; L: 光照。"

图6

Top50的DMs (顶部)与Top100的DEGs (左侧)之间的相关性热图(a)以及代谢-转录KEGG共有富集通路气泡图(b) (a) 红色代表正相关, 蓝色代表负相关。椭圆越扁, 代表相关性的绝对值越高。相关性统计检验P < 0.05会进行星号(*)标记。(b) 三角形代表差异基因, 圆点代表差异代谢物。-log10(P-value)越大, 富集可信度越高。S: 遮阴; L: 光照。"

图7

苯丙素合成途径DMs的含量热图(a)以及DMs与DEGs之间的相关性热图(b)"

图8

糖代谢相关途径DMs的含量热图(a)以及DMs与DEGs之间的相关性热图(b)"

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