1.Differential Analysis of Flavonoid-regulated Genes in Two Cultivars of Angelica sinensis Based on Hybrid Sequencing Strategy
Li XU ; Tiantian ZHU ; Ling JIN ; Shuqi KANG ; Minghui ZHANG ; Yuanyuan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(1):141-147
ObjectiveTo compare the transcriptional levels of two cultivars (Mingui 1 and Mingui 2) with different stem and leaf colors. MethodThe fresh leaves with petioles and the upper stems from Angelica sinensis with two colors were selected as the material, and the hybrid sequencing strategy was used. The technology at the transcriptome was used to build the non-reference full-length transcript library of A. sinensis, and the RNA-seq technology was used to analyze differentially expressed genes of the two cultivars, reusing a public database for biology function annotation and fine classification of differentially expressed genes. The main candidate genes regulating color differences between stems and leaves of A. sinensis were screened out. ResultThe sequencing results of A. sinensis transcripts were good and the quality of the sequencing data was high. The 34 528 full-length transcripts were annotated into 33 947, 33 241, 29 150, and 22 601 in the Non-redundant Protein Sequence Database (NR), Kyoto Encyclopedia of Genes and Genomes (KEGG), SwissProt, and Clusters of Orthologous Groups for Eukaryotic Complete Genomes (KOG), respectively. The 705 differentially expressed genes of the two cultivars with biological and molecular functions were divided into 11 categories, which were mainly enriched in the primary metabolism (17.87%), stress response (14.47%), and secondary metabolism (11.49%). The differentially expressed genes related to colors were mainly concentrated in the flavonoid biosynthesis pathway. ConclusionThe main reason for the color differences of the stems and leaves in two cultivars of A. sinensis may be related to the expression differences of genes regulating flavonoid biosynthesis, which lays a foundation for subsequent functional verification and further clarification of the relationship with the main pharmacodynamic components of A. sinensis.
2.Identification and Genetic Variation Analysis of Wild and Cultivated Germplasm of Angelica sinensis Based on cp DNA
Minghui ZHANG ; Tiantian ZHU ; Ling JIN ; Fusheng WANG ; Shuqi KANG ; Li XU ; Yuanyuan WANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(15):129-138
ObjectiveTo conduct genetic variation analysis of 11 cultivars and 7 wild populations of Angelica sinensis in Gansu province based on the chloroplast gene (cp DNA), and provide references for germplasm identification and breeding of new cultivars of A. sinensis. MethodThree pairs of cp DNA primers were used for polymerase chain reaction (PCR) amplification and sequencing of A. sinensis samples. MegaX was used to perform statistics on sequence characteristics and calculate mean genetic distances among A. sinensis populations. Unweighted pair-group method with arithmetic means (UPGMA) clustering tree based on genetic distance was constructed by NTSYS 2.10e. DanSP v6 was used to calculate sequence polymorphism and Tajima's D of A. sinensis. PERMUT was used to calculate the population structure of A. sinensis. Arlequin v3.5 was used to perform molecular variation analysis, and PopART1.7 was used to construct TCS haplotype network. ResultThree pairs of cp DNA primers were amplified, sequenced, compared, and combined to give a sequence length of 1 759 bp. One variable site was detected in the wild A. sinensis and 480 variable sites were detected in the cultivated A. sinensis, including 97 singleton variable sites, 383 parsimony informative sites, and 152 insertion-deletion sites. In the three regions of matK, psbA-trnH, and rbcL of cp DNA in the wild and cultivated A. sinensis, matK was the region with the highest polymorphism. Tajima’s D of all the combined sequences of A. sinensis were not significantly negative, but psbA-trnH and rbcL genes of the cultivated A. sinensis were significantly negative, indicating that the A. sinensis followed neutral evolution on a whole, while psbA-trnH and rbcL genes had undergone selection. The degree of genetic differentiation (Fst=0) among wild populations was lower than that among cultivated populations (Fst=0.114 19, P<0.05). The degree of genetic differentiation between wild and cultivated A. sinensis was relatively high (Fst=0.942 55, P<0.01). Genetic variation in the cultivated A. sinensis was mainly found within the populations (89%). UPGMA cluster tree based on genetic distance showed that the wild A. sinensis and the cultivated A. sinensis were clustered into one branch, respectively, with a distant genetic relationship, and the population 3 in the cultivated A. sinensis was far from other cultivated populations. The TCS haplotype network consisted of 15 haplotypes and 4 unknown haplotypes, which was divided into 3 parts, with a large number of variations among each part. Shared haplotypes were only found in the wild or cultivated groups, and there were no shared haplotypes between groups. ConclusionThe genetic diversity of A. sinensis was low at species level, and the population diversity of the wild was lower than that of the cultivated. The degree of genetic differentiation between the wild and the cultivated A. sinensis was high, but that in the wild and the cultivated populations were low. Genetic variation in the cultivated A. sinensis was mainly found within populations.
3.Genetic Diversity Analysis of Isatis indigotica Germplasm Materials by Chloroplast DNA Variation and ISSR Molecular Marker
Tiantian ZHU ; Tao DU ; Ling JIN ; Fusheng WANG ; Shuqi KANG ; Li XU ; Minghui ZHANG ; He LIN
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(18):117-126
ObjectiveTo analyze the sequence variation and genetic diversity of 47 Isatis indigotica germplasm materials, and carry out the study on the genetic differentiation and structure. MethodGenomic DNA of 47 I. indigotica germplasm materials were extracted by kit extraction method. Two chloroplast DNA (cp DNA) sequences and five inter-simple sequence repeat (ISSR) primers were used for amplification and sequencing. Chromas, Mega 7.0, DanSP5, and GenALEx were used to calibrate, splice, and analyze the sequence characteristics. PERMUT and PopGen 1.31 were used to analyze the genetic diversity parameters and genetic structure, and NTSYS was used to obtain the unweighted pair-group method with arithmetic means(UPGMA) clustering tree plot of 47 I. indigotica germplasm materials. ResultA total of 129 samples from 47 I. indigotica germplasm materials were successfully amplified and sequenced. The length of 2 cp DNA sequences after spliced was 1 412 bp, and there were 377 polymorphic variation loci, and 36 haplotypes. Fu and Li's D* test was significant (P<0.01). The values of Pi, HS, and HT based on cp DNA were 0.119 89, 0.787, and 0.891, respectively. The genetic differentiation coefficients of gene differentiation coefficient(Gst), nucleotide differentiation coefficient(Nst), and fixation index(Fst) were 0.117, 0.468, and 0.488, respectively, and the gene flow (Nm) was 0.615. The mean values of PPB, Shannon information diversity index(I), Nei's genetic diversity index(H), and Gst based on ISSR were 78.85%, 0.334 8, 0.218 6, and 0.754 4, respectively, and the Nm value was 0.162 8. ConclusionI. indigotica has high genetic diversity and abundant haplotypes at the species level, with abundant haplotypes. Genetic differentiation among different germplasm materials is obvious, and gene exchange is not frequent. Genetic variation mainly exists among populations. The population has accumulated various low-frequency gene mutations recently, suggesting that it has experienced significant regional expansion in the history.
4.High-quality Formation of Cistanches Herba: A Review
Jing ZHANG ; Tiantian ZHU ; Ling JIN ; Tianle LIU ; Minghui ZHANG ; Li XU ; Shuqi KANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(18):259-266
With the continuous development of traditional Chinese medicine (TCM), the requirement for the quality of Chinese medicines has become increasingly higher since they have been widely used in clinical practice. Chinese medicinal materials are the material basis for the inheritance and development of TCM, and their quality directly affects the clinical efficacy. Studying the quality of Chinese medicinal materials is the key to ensure the quality and realize the large-scale application. As one of rare Chinese medicinal materials, Cistanches Herba has the functions of tonifying kidney yang, invigorating blood and essence, moistening intestines to relieve constipation. High-quality Cistanches Herba is characterized by glossy appearance, high density, fleshy and soft texture, and sweet taste. With the reduction of wild resources, the products from cultivated Cistanche deserticola or C. tubulosa become dominant on the market of Cistanches Herba. The cultivation areas are widely distributed, mainly concentrated in Gansu, Inner Mongolia, and Xinjiang. However, the cultivated products have varied quality due to the differences in germplasm, producing region, cultivation method, harvesting, and processing. According to the theories of quality evaluation based on morphological characteristics and excellent appearance indicating high quality, this paper reviewed the literature on the quality evaluation, growth, development, and processing of Cistanches Herba in the last decade to explore the main factors (genetic characteristics, environmental conditions, and harvesting and processing factors) affecting the quality of Cistanches Herba. The review aims to explore the factors for the high quality and provide a reference for the producing region screening, directional cultivation, and production of Cistanches Herba.
5.Genomic and transcriptomic analysis unveils population evolution and development of pesticide resistance in fall armyworm Spodoptera frugiperda.
Furong GUI ; Tianming LAN ; Yue ZHAO ; Wei GUO ; Yang DONG ; Dongming FANG ; Huan LIU ; Haimeng LI ; Hongli WANG ; Ruoshi HAO ; Xiaofang CHENG ; Yahong LI ; Pengcheng YANG ; Sunil Kumar SAHU ; Yaping CHEN ; Le CHENG ; Shuqi HE ; Ping LIU ; Guangyi FAN ; Haorong LU ; Guohai HU ; Wei DONG ; Bin CHEN ; Yuan JIANG ; Yongwei ZHANG ; Hanhong XU ; Fei LIN ; Bernard SLIPPERS ; Alisa POSTMA ; Matthew JACKSON ; Birhan Addisie ABATE ; Kassahun TESFAYE ; Aschalew Lemma DEMIE ; Meseret Destaw BAYELEYGNE ; Dawit Tesfaye DEGEFU ; Feng CHEN ; Paul K KURIA ; Zachary M KINYUA ; Tong-Xian LIU ; Huanming YANG ; Fangneng HUANG ; Xin LIU ; Jun SHENG ; Le KANG
Protein & Cell 2022;13(7):513-531
The fall armyworm (FAW), Spodoptera frugiperda, is a destructive pest native to America and has recently become an invasive insect pest in China. Because of its rapid spread and great risks in China, understanding of FAW genetic background and pesticide resistance is urgent and essential to develop effective management strategies. Here, we assembled a chromosome-level genome of a male FAW (SFynMstLFR) and compared re-sequencing results of the populations from America, Africa, and China. Strain identification of 163 individuals collected from America, Africa and China showed that both C and R strains were found in the American populations, while only C strain was found in the Chinese and African populations. Moreover, population genomics analysis showed that populations from Africa and China have close relationship with significantly genetic differentiation from American populations. Taken together, FAWs invaded into China were most likely originated from Africa. Comparative genomics analysis displayed that the cytochrome p450 gene family is extremely expanded to 425 members in FAW, of which 283 genes are specific to FAW. Treatments of Chinese populations with twenty-three pesticides showed the variant patterns of transcriptome profiles, and several detoxification genes such as AOX, UGT and GST specially responded to the pesticides. These findings will be useful in developing effective strategies for management of FAW in China and other invaded areas.
Animals
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China
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Genomics
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Humans
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Male
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Pesticides
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Spodoptera/genetics*
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Transcriptome