





中国农业科学 ›› 2019, Vol. 52 ›› Issue (22): 4002-4015.doi: 10.3864/j.issn.0578-1752.2019.22.006
收稿日期:2019-06-14
接受日期:2019-08-12
出版日期:2019-11-16
发布日期:2019-11-16
联系方式:
张飞,Tel:024-31029903;E-mail:zhangfei19821121@163.com
基金资助:
ZHANG Fei,WANG YanQiu,ZHU Kai,ZHANG ZhiPeng,ZHU ZhenXing,LU Feng(
),ZOU JianQiu(
)
Received:2019-06-14
Accepted:2019-08-12
Published:2019-11-16
Online:2019-11-16
摘要:
【目的】土壤盐渍化是制约作物生产的重要非生物胁迫因子之一,高粱耐盐性强,进行高粱耐盐基因挖掘及分子机制研究是开发和利用盐渍土壤的有效途径,通过转录组测序分析与高粱耐盐相关的基因调控机制和代谢通路,挖掘高粱耐盐潜力。【方法】 通过以筛选出的极耐盐品种八叶齐和盐极敏感品种PL212为试验材料,采用盆栽沙培,在播种后20 d(5叶期)采用180 mmol·L -1的 NaCl 溶液漫灌模拟盐逆境,盐胁迫48 h后取幼叶,并连同对照(未经过盐处理)的同期幼苗共4个样品提取RNA,进行转录组测序,采用qRT-PCR方法对测序结果进行验证。 【结果】 耐盐和盐敏感材料分别在盐渍和非盐渍处理下的4个样品间共检测到1 338个差异表达基因,包括819个上调基因和519个下调基因。聚类分析发现在应答盐渍胁迫逆境时,5个依赖性氧合酶超家族蛋白、4个富含半胱氨酸的激酶、3个谷胱甘肽S-转移酶和3个重金属运输/解毒超家族蛋白相关基因表现出明显的上调表达和下调表达,还发现1个K +转运蛋白基因在耐盐调节中起着重要作用。GO分析发现在15 418个基因中获得4 528个有效GO注释条目,同时耐盐和盐敏感材料在遭受盐逆境时的生物过程、细胞组分和分子功能3个方面均存在较大差异。生物过程中代谢过程、细胞过程耐盐材料明显高于盐敏感材料,耐盐材料的生理过程中较盐敏感材料增加了多生物过程和定位这两个过程,很可能与耐盐材料盐抗性较强密切相关。差异基因KEGG分析结果显示耐盐和盐敏感材料在对照和盐渍胁迫条件下的苯丙烷类生物合成、苯丙氨酸代谢、类黄酮生物合成3个途径中差异基因表达较多,可能是造成耐盐和盐敏感材料耐盐性差异较大的重要原因。 【结论】 高粱耐盐调控基因表达涉及生物过程、细胞组分和分子功能多个方面,生物过程和定位这两个过程是提高高粱耐盐性的关键;苯丙烷类生物合成、苯丙氨酸代谢、类黄酮生物合成3个途径的基因表达很可能是造成盐害的重要原因。
张飞,王艳秋,朱凯,张志鹏,朱振兴,卢峰,邹剑秋. 不同耐盐性高粱在盐逆境下的比较转录组分析[J]. 中国农业科学, 2019, 52(22): 4002-4015.
ZHANG Fei,WANG YanQiu,ZHU Kai,ZHANG ZhiPeng,ZHU ZhenXing,LU Feng,ZOU JianQiu. Comparative Transcriptome Analysis of Different Salt Tolerance Sorghum (Sorghum bicolor L. Moench) Under Salt Stress[J]. Scientia Agricultura Sinica, 2019, 52(22): 4002-4015.
表1
测序数据质量统计"
| 样品 Sample | 序列长度 Length (bp) | Bases (Billion) | Q20比例 Q20 percentage (%) | Q30比例 Q30 percentage (%) | G和C占总碱基数量百分比 GC(%) |
|---|---|---|---|---|---|
| CK-耐盐 CK-tolerant | 147.45 | 64.89 | 96.09 | 90.20 | 52.18 |
| CK-盐敏感 CK-sensitive | 147.57 | 61.35 | 95.85 | 89.74 | 53.44 |
| Salt-耐盐 Salt-tolerant | 147.64 | 61.72 | 95.85 | 89.73 | 52.38 |
| Salt-盐敏感 Salt-sensitive | 147.41 | 61.95 | 95.93 | 89.89 | 53.28 |
表2
不同表达水平区间的基因数量及比例统计"
| 项目 Item | 样品 Sample | RPKM值RPKM value | |||||
|---|---|---|---|---|---|---|---|
| 0—0.1 | 0.1—1 | 1—3 | 3—15 | 15—60 | >60 | ||
| 基因数量 Number of genes | CK-耐盐CK-tolerant | 2125 | 6815 | 4570 | 8046 | 4345 | 1371 |
| CK-盐敏感CK-sensitive | 2392 | 6985 | 4621 | 9605 | 3890 | 1377 | |
| Salt-耐盐Salt-tolerant | 2075 | 6687 | 4537 | 8024 | 4436 | 1364 | |
| Salt-盐敏感Salt-sensitive | 2258 | 6864 | 4625 | 7811 | 3927 | 1281 | |
| 基因表达比例 Gene expression ratio (%) | CK-耐盐CK-tolerant | 7.79 | 24.98 | 16.75 | 29.49 | 15.96 | 5.03 |
| CK-盐敏感CK-sensitive | 8.90 | 26.00 | 17.20 | 28.30 | 14.48 | 5.03 | |
| Salt-耐盐Salt-tolerant | 7.65 | 24.65 | 16.73 | 29.58 | 16.36 | 5.03 | |
| Salt-盐敏感Salt-sensitive | 8.44 | 25.64 | 17.28 | 29.18 | 14.67 | 4.79 | |
表3
盐胁迫下4个高粱样品间的差异表达基因"
| 样品比较 Sample-VS-Sample | 上调基因 Up-regulated genes | 下调基因 Down-regulated genes |
|---|---|---|
| CK-耐盐VS CK-盐敏感 CK-tolerant VS CK-sensitive | 346 | 134 |
| Salt-耐盐VS Salt-盐敏感 Salt-tolerant VS Salt-sensitive | 353 | 200 |
| Salt-耐盐VS CK-耐盐 Salt-tolerant VS CK-tolerant | 62 | 122 |
| Salt-盐敏感VS CK-盐敏感 Salt-sensitive VS CK-sensitive | 58 | 63 |
表4
高粱耐盐相关基因及功能描述"
| 基因ID Gene_ID | log2FC | KOG | KEGG/ec | Best-hit-Arabi-name | Arabi-symbol | 功能描述 Arabi-decline |
|---|---|---|---|---|---|---|
| Sobic.001G314300.v3.1 | 2.29 | KOG0143 | 1.14.11.9 | AT5G24530.1 | DMR6 | 铁离子转运蛋白 Fe ion transport protein |
| Sobic.006G190000.v3.1 | 1.91 | KOG0143 | 1.14.11.9 | AT5G24530.1 | DMR6 | 铁离子转运蛋白 Fe ion transport protein |
| Sobic.001G526900.v3.1 | 1.40 | KOG0143 | 1.14.11.9 | AT5G24530.1 | DMR6 | 铁离子转运蛋白 Fe ion transport protein |
| Sobic.001G166401.v3.1 | 1.27 | KOG0143 | AT3G19000.1 | 铁离子转运蛋白 Fe ion transport protein | ||
| Sobic.009G044400.v3.1 | 1.19 | KOG0143 | 1.14.17.4 | AT1G77330.1 | 铁离子转运蛋白 Fe ion transport protein | |
| Sobic.001G215900.v3.1 | 1.29 | KOG1187 | 2.7.11.1 | AT1G70520.1 | CRK2 | 富含半胱氨酸的蛋白激酶 Cysteine-rich protein kinase |
| Sobic.002G327700.v3.1 | 1.18 | KOG1187 | 2.7.11.1 | AT4G23310.1 | CRK23 | 富含半胱氨酸的蛋白激酶 Cysteine-rich protein kinase |
| Sobic.002G327800.v3.1 | 1.23 | KOG1187 | 2.7.11.1 | AT4G05200.1 | CRK25 | 富含半胱氨酸的蛋白激酶 Cysteine-rich protein kinase |
| Sobic.008G099300.v3.1 | 1.72 | 2.7.11.1 | AT4G00970.1 | CRK41 | 富含半胱氨酸的蛋白激酶 Cysteine-rich protein kinase | |
| Sobic.003G164800.v3.1 | 1.33 | KOG0867 | 2.5.1.18 | AT3G62760.1 | ATGSTF13 | 谷胱甘肽S-转移酶家族蛋白 Glutathione S-transferase family protein |
| Sobic.001G318900.v3.1 | 2.12 | KOG0406 | 2.5.1.18 | AT1G10360.1 | ATGSTU18,GST29, GSTU18 | 谷胱甘肽S-转移酶TAU 18 Glutathione S-transferase TAU 18 |
| Sobic.001G318200.v3.1 | 2.02 | KOG0406 | 2.5.1.18 | AT1G10360.1 | ATGSTU18,GST29,GSTU18 | 谷胱甘肽S-转移酶TAU 18 Glutathione S-transferase TAU 18 |
| Sobic.006G113800.v3.1 | 2.24 | AT5G48290.1 | 重金属运输/解毒超家族蛋白 Heavy metal transport/detoxification superfamily protein | |||
| Sobic.006G114200.v3.1 | 1.93 | AT5G48290.1 | 重金属运输/解毒超家族蛋白 Heavy metal transport/detoxification superfamily protein | |||
| Sobic.006G257700.v3.1 | 1.53 | KOG1603 | AT1G01490.1 | 重金属运输/解毒超家族蛋白 Heavy metal transport/detoxification superfamily protein | ||
| Sobic.002G220600.v3.1 | 1.09 | AT2G30070.1 | ATKT1,ATKT1P,ATKUP1,KT1,KUP1 | 钾转运蛋白 Potassium transport protein | ||
| Sobic.002G416600.v3.1 | 1.97 | 1.11.1.7 | 过氧化物酶超家族蛋白 Peroxidase superfamily protein |
表5
高粱耐盐基因分布及注释到基因"
| 基因本体 Gene ontology term | 集群频率 (出现集群/总集群数量) Cluster frequency(Appears number/total number) | 基因组使用频率 Genome frequency of use | 修正P值 Corrected P-value | 注释到基因集群 Annotated gene cluster |
|---|---|---|---|---|
| 膜 Membrane | 12 /14 85.7% | 1862/15418, 12.1% | 5.37e-09 | Sobic.004G073400.v3.1, Sobic.002G329600.v3.1, Sobic.007G210500.v3.1, Sobic.004G182300.v3.1, Sobic.006G003700.v3.1, Sobic.002G339100.v3.1, Sobic.002G367700.v3.1, Sobic.005G037300.v3.1, Sobic.002G220600.v3.1, Sobic.006G021900.v3.1, Sobic.002G201900.v3.1, Sobic.010G146100.v3.1 |
| 膜部分 Membrane part | 9/14 64.3% | 1043 /15418 6.8% | 3.37e-07 | Sobic.002G201900.v3.1, Sobic.006G021900.v3.1, Sobic.005G037300.v3.1, Sobic.004G182300.v3.1, Sobic.007G210500.v3.1, Sobic.002G329600.v3.1, Sobic.004G073400.v3.1, Sobic.002G339100.v3.1, Sobic.006G003700.v3.1 |
| 膜整体 Integral to membrane | 8 /14 57.1% | 829 /15418 5.4% | 1.21e-06 | Sobic.004G182300.v3.1, Sobic.004G073400.v3.1, Sobic.007G210500.v3.1, Sobic.002G339100.v3.1, Sobic.006G003700.v3.1, Sobic.006G021900.v3.1, Sobic.002G201900.v3.1, Sobic.005G037300.v3.1 |
| 膜固有 Intrinsic to membrane | 8 /14 57.1% | 841 /15418 5.5% | 1.35e-06 | Sobic.002G201900.v3.1, Sobic.006G021900.v3.1, Sobic.005G037300.v3.1, Sobic.004G182300.v3.1, Sobic.007G210500.v3.1, Sobic.004G073400.v3.1, Sobic.002G339100.v3.1, Sobic.006G003700.v3.1 |
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