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通过从头组装对青藏高原菊芋(L.)转录组进行表征,以发现与果聚糖合成相关的基因并进行SSR分析。

Characterization of the Tibet plateau Jerusalem artichoke ( L.) transcriptome by de novo assembly to discover genes associated with fructan synthesis and SSR analysis.

作者信息

Yang Shipeng, Sun Xuemei, Jiang Xiaoting, Wang Lihui, Tian Jie, Li Li, Zhao Mengliang, Zhong Qiwen

机构信息

1Academy of Agriculture and Forestry Sciences of Qinghai University (Qinghai Academy of Agriculture and Forestry Sciences), Qinghai Key Laboratory of Vegetable Genetics and Physiology, State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining, 810016 China.

Qinghai Higher Vocational & Technical College Institute, Ledu, 810799 China.

出版信息

Hereditas. 2019 Feb 6;156:9. doi: 10.1186/s41065-019-0086-8. eCollection 2019.

Abstract

BACKGROUND

Jerusalem artichoke ( L.) is a characteristic crop in the Qinghai-Tibet Plateau which has rapidly developed and gained socioeconomic importance in recent years. Fructans are abundant in tubers and represent the foundation for their formation, processing and utilization of yield; and are also widely used in new sugar-based materials, bioenergy processing, ecological management, and functional feed. To identify key genes in the metabolic pathway of fructans in Jerusalem artichoke, high-throughput sequencing was performed using Illumina Hi Seq™ 2500 equipment to construct a transcriptome library.

RESULTS

Qinghai-Tibet Plateau Jerusalem artichoke "Qingyu No.1" was used as the material; roots, stems, leaves, flowers and tubers of Jerusalem artichoke in its flowering stage were mixed into a mosaic of the Jerusalem artichoke transcriptome library, obtaining 63,089 unigenes with an average length of 713.6 bp. Gene annotation through the Nr, Swiss Prot, GO, KOG and KEGG databases revealed 34.95 and 46.91% of these unigenes had similar sequences in the Nr and Swiss Prot databases. The GO classification showed the Jerusalem artichoke unigenes were divided into three ontologies, with a total of 49 functional groups encompassing biological processes, cellular components, and molecular functions. Among them, there were more unigenes involved in the functional groups for cellular processes, metabolic processes, and single-organism processes. 38,999 unigenes were annotated by KOG and divided into 25 categories according to their functions; the most common annotation being general function prediction. A total of 13,878 unigenes (22%) were annotated in the KEGG database, with the largest proportion corresponding to pathways related to carbohydrate metabolism. A total of 12 unigenes were involved in the synthesis and degradation of fructan. Cluster analysis revealed the candidate 12 unigene proteins were dispersed in the 5 major families of proteins involved in fructan synthesis and degradation. The synergistic effect of INV gene is necessary during fructose synthesis and degradation in Jerusalem artichoke tuber development. The sequencing data from the transcriptome of this species can provide a reliable data basis for the identification and assessment of the expression of the members of the INV gene family.A simple sequence repeat (SSR) loci search was performed on the transcriptome data of Jerusalem artichoke, identifying 6635 eligible SSR loci with a large proportion of dinucleotide and trinucleotide repeats, and the most different motifs were repeated 5 times and 6 times. Dinucleotide and trinucleotide repeat motifs were the most frequent, with AG/CT and ACC/GGT repeat motifs accounting for the highest proportion.

CONCLUSIONS

In this study, a database search of the transcriptome of the Jerusalem artichoke from the Qinghai Tibet Plateau was conducted by high throughput sequencing technology to obtain important transcriptional and SSR loci information. This allowed characterization of the overall expression features of the Jerusalem artichoke transcriptome, identifying the key genes involved in metabolism in this species. In turn, this offers a foundation for further research on the regulatory mechanisms of fructan metabolism in Jerusalem artichoke.

摘要

背景

菊芋是青藏高原的特色作物,近年来发展迅速并具有重要的社会经济价值。菊糖在块茎中含量丰富,是其产量形成、加工利用的基础;并且在新型糖基材料、生物能源加工、生态治理及功能性饲料等方面也有广泛应用。为鉴定菊芋中菊糖代谢途径的关键基因,利用Illumina Hi Seq™ 2500设备进行高通量测序以构建转录组文库。

结果

以青藏高原菊芋“青芋1号”为材料,取其花期的根、茎、叶、花和块茎混合构建菊芋转录组文库,获得63,089条单基因,平均长度为713.6 bp。通过Nr、Swiss Prot、GO、KOG和KEGG数据库进行基因注释,结果显示这些单基因中分别有34.95%和46.91%在Nr和Swiss Prot数据库中有相似序列。GO分类表明菊芋单基因被分为三个本体,共有49个功能组,涵盖生物过程、细胞组分和分子功能。其中,参与细胞过程、代谢过程和单细胞过程功能组的单基因较多。38,999条单基因经KOG注释并根据功能分为25类,最常见的注释为一般功能预测。共有13,878条单基因(22%)在KEGG数据库中得到注释,其中比例最大的对应于与碳水化合物代谢相关的途径。共有12条单基因参与菊糖的合成与降解。聚类分析表明候选的12个单基因蛋白分散在参与菊糖合成与降解的5个主要蛋白家族中。在菊芋块茎发育过程中,INV基因的协同作用在果糖合成与降解过程中是必需的。该物种转录组的测序数据可为INV基因家族成员的鉴定及表达评估提供可靠的数据基础。对菊芋转录组数据进行简单序列重复(SSR)位点搜索,鉴定出6635个合格的SSR位点,其中二核苷酸和三核苷酸重复比例较大,最不同的基序分别重复5次和6次。二核苷酸和三核苷酸重复基序最为常见,AG/CT和ACC/GGT重复基序占比最高。

结论

本研究通过高通量测序技术对青藏高原菊芋转录组进行数据库搜索,获得重要的转录和SSR位点信息。这使得能够表征菊芋转录组的整体表达特征,鉴定该物种中参与代谢的关键基因。进而为进一步研究菊芋中菊糖代谢的调控机制提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7990/6364414/ef6278427808/41065_2019_86_Fig1_HTML.jpg

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