五种不同组织结构特点昆虫的总RNA提取效率比较 *

doi:10.7679/j.issn.2095-1353.2020.102

应用昆虫学报 ›› 2020, Vol. 57 ›› Issue (4): 988-994.doi: 10.7679/j.issn.2095-1353.2020.102

• 技术与方法 • 上一篇

五种不同组织结构特点昆虫的总RNA提取效率比较 ^*

尹志勇¹(), 郭建军¹, 陈绪美¹, 郑玉琳¹, 田莹¹, 檀军¹^,²^,^***()

1. 贵州大学昆虫研究所,贵州山地农业病虫害重点实验室,贵阳 550025
2. 遵义医科大学,组织学与胚胎学教研室,遵义 563003

收稿日期:2020-01-06 接受日期:2020-04-15 出版日期:2020-07-27 发布日期:2020-09-02
通讯作者: 檀军 E-mail:2329723038@qq.com;juntan@zmu.edu.cn
基金资助:
*国家自然科学基金项目(81803968);资源昆虫学研究科技创新团队(黔教合人才团队字[2015]71);贵州省昆虫信息系统与资源开发利用重点实验室(黔科合LH字[2015]7685号)

Comparison of the effectiveness of the Trizol method versus an RNA extraction kit for extracting total RNA from insect species with different tissue characteristics

Zhi-Yong YIN¹(), Jian-Jun GUO¹, Xu-Mei CHEN¹, Yu-Lin ZHENG¹, Ying TIAN¹, Jun TAN¹^,²^,^***()

1. Institute of Entomology, Guizhou University, Guizhou Provincial Key Laboratory for Agricultural Pest Management of the Mountainous Region, Guiyang 550025, China
2. Department of Histology and Embryology, Zunyi Medical University, Zunyi 563000, China

Received:2020-01-06 Accepted:2020-04-15 Online:2020-07-27 Published:2020-09-02
Contact: Jun TAN E-mail:2329723038@qq.com;juntan@zmu.edu.cn

摘要/Abstract

摘要：

【目的】 比较RNAeasy抽提试剂盒法和Trizol提取法对不同种昆虫总RNA提取的效率,为不同种昆虫总RNA提取方法选择提供参考。【方法】 采用RNAeasy抽提试剂盒法和Trizol提取法分别对九香虫Aspongopus chinensis Dallas、白背飞虱Sogatella furcifera Horváth、中华蜜蜂Apis cerana cerana、黄蜻Pantala flavescens和蒿金叶甲Chrysolina aurichalcea 进行总RNA提取,通过紫外分光光度计测定总RNA吸光度（OD）值,利用琼脂糖凝胶电泳检测等方式对RNA质量进行评估。【结果】2种提取方法提取不同昆虫总RNA中,Trizol法提取白背飞虱（同翅目）、蒿金叶甲（鞘翅目）和九香虫（半翅目）的总RNA浓度较高,但纯度较低。电泳结果显示总RNA有降解,完整性较差,RNAeasy抽提试剂盒法提取黄蜻（蜻蜓目）和中华蜜蜂（膜翅目）的浓度较低,纯度较高,电泳结果显示完整性较好。【结论】 Trizol法更适合提取小型或几丁质含量高的昆虫,RNA抽提试剂盒法更适合几丁质含量较少、腹软且有大量体液的昆虫总RNA提取。

关键词: 昆虫总RNA, 提取方法, 提取效率, 紫外分光光度, 琼脂糖凝胶电泳

Abstract:

[Objectives] The relative efficiency of using a RNA extraction kit to extract total RNA from different insects was compared to that of the Trizol method. [Methods] An RNA extraction kit and Trizol were used to extract total RNA from five insects; Aspongopus chinensis Dallas, Sogatella furcifera Horváth, Apis cerana cerana, Pantala flavescens and Chrysolina aurichalcea. The quality and the OD value of total RNA obtained were measured with a UV spectrophotometer and agarose gel electrophoresis. [Results] Although a higher concentration of total RNA was extracted with the Trizol method from three species, S. furcifera, C. aurichalcea, and A. chinensis, it had lower purity and electrophoresis results indicated that it was degraded with poor integrity. Conversely, although the concentration of total RNA obtained with an RNA extraction kit from P. flavescens (Odonata) and A. cerana cerana (Hymenoptera) was relatively low, it was relatively pure and had better integrity. [Conclusion] The trizol method is more suitable for extracting total RNA from small insects or those with high chitin content, whereas an RNA extraction kit is better for insects with low chitin content, a soft abdomen and more body fluid.

Key words: insect RNA, extraction method, extraction efficiency, ultraviolet spectrophotometer, agarose gel electrophoresis

尹志勇, 郭建军, 陈绪美, 郑玉琳, 田莹, 檀军. 五种不同组织结构特点昆虫的总RNA提取效率比较 ^*[J]. 应用昆虫学报, 2020, 57(4): 988-994.

Zhi-Yong YIN, Jian-Jun GUO, Xu-Mei CHEN, Yu-Lin ZHENG, Ying TIAN, Jun TAN. Comparison of the effectiveness of the Trizol method versus an RNA extraction kit for extracting total RNA from insect species with different tissue characteristics[J]. Chinese Journal of Applied Entomology, 2020, 57(4): 988-994.

图/表 5

表1 单只昆虫质量（g）

Table 1 Insect mass and eluent volume

	Trizol法 Trizol method	RNAeasy抽提试剂盒法 RNAeasy extraction kit method
白背飞虱 Sogatella furcifera Horváth	0.001	0.001
蒿金叶甲 Chrysolina aurichalcea	0.024	0.024
黄蜻 Pantala flavescens	0.206	0.261
中华蜜蜂 Apis cerana cerana Fabricius	0.093	0.084
九香虫 Aspongopus chinensis Dallas	0.387	0.384

表2 2种RNA提取方法提取的RNA含量（ng/mg）

Table 2 RNA content extracted by two RNA extraction methods (ng/mg)

	白背飞虱 Sogatella furcifera	蒿金叶甲 Chrysolina aurichalcea	黄蜻 Pantala flavescens	中华蜜蜂 Apis cerana cerana	九香虫 Aspongopus chinensis
Trizol法 Trizol method	8 868.33±3 758.77^*	2 058.47±1 398.60^*	482.48±258.13	145.65±47.15	147.86±28.19
RNAeasy抽提试剂盒法 RNAeasy extraction kit method	213.33±10.14	156.11±41.22	76.30±3.28	93.33±7.56	63.16±17.32

表3 2种RNA提取方法提取的OD260/280

Table 3 OD260/280 of RNA extracted by two methods

	白背飞虱 Sogatella furcifera	蒿金叶甲 Chrysolina aurichalcea	黄蜻 Pantala flavescens	中华蜜蜂 Apis cerana cerana	九香虫 Aspongopus chinensis
Trizol法Trizol method	1.54±0.02^**	1.90±0.11	1.88±0.03^**	1.68±0.01^**	1.79±0.06^**
RNAeasy抽提试剂盒法RNAeasy extraction kit method	2.02±0.04	2.19±0.01	2.15±0.003	2.09±0.01	2.06±0.01

图1 琼脂糖凝胶电泳检测Trizol法提取昆虫总RNA图 A. 九香虫;B. 中华蜜蜂;C. 黄蜻;D. 蒿金叶甲;E. 白背飞虱;泳道1-3,4-6,7-9代表3个重复实验结果。

Fig. 1 Agarose gel electrophoresis detect the RNA extracted by Trizol method A. A. chinensis; B. A. cerana cerana; C. P. flavescens; D. C. aurichalcea; E. S. furcifera; Lane 1-3, 4-6, 7-9 represent 3 independent experiments.

图2 RNAeasy动物RNA抽提试剂盒法琼脂糖凝胶电泳图 A. 九香虫;B. 中华蜜蜂;C. 黄蜻;D. 蒿金叶甲;E. 白背飞虱;泳道1-3,4-6,7-9代表3个重复实验结果。

Fig. 2 Agarose gel electrophoresis by RNAeasy animal RNA extraction kit A. A. chinensis; B. A. cerana cerana; C. P. flavescens; D. C. aurichalcea; E. S. furcifera; Lane 1-3, 4-6, 7-9 represent 3 independent experiments.

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五种不同组织结构特点昆虫的总RNA提取效率比较 ^*

Comparison of the effectiveness of the Trizol method versus an RNA extraction kit for extracting total RNA from insect species with different tissue characteristics

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