molecular marker

分子标记
  • 文章类型: Journal Article
    盐碱地的利用是全球性的挑战,培育耐盐大豆品种有利于提高盐碱地利用效率。探索耐盐大豆品种的遗传基础,开发耐盐分子标记,可有效促进大豆耐盐育种进程。在研究中,采用隶属函数法对大豆苗期与耐盐性和综合耐盐性相关的7个性状进行评价;对含有200份大豆材料的自然群体进行全基因组关联分析(GWAS);对112个重组自交系(RIL)群体进行连锁分析,检测耐盐性的数量性状位点(QTLs)。在GWAS中,绘制了147个SNP,解释5.28-17.16%的表型变异。在联系分析中,确定了10个QTL,这可以解释6.9-16.16%的表型变异。结果发现,自然种群和RIL种群之间存在两个共同定位的区域,含有7个大豆耐盐候选基因。此外,发现一个共定位区间包含qZJS-15-1,rs47665107和rs4793412,所有这些都可以解释超过10%的表型变异率,使其适合分子标记开发。rs47665107和rs47934112的物理位置包含在qZJS-15-1中。因此,使用Chr设计和开发了KASP标记。15:47907445,与qZJS-15-1密切相关。该标记可以准确,清晰地将所测试的杂合群体中耐盐基因型的材料聚类。研究发现的QTL和KASP标记为加快大豆耐盐育种提供了理论和技术依据。
    The utilization of saline land is a global challenge, and cultivating salt-tolerant soybean varieties is beneficial for improving the efficiency of saline land utilization. Exploring the genetic basis of salt-tolerant soybean varieties and developing salt-tolerant molecular markers can effectively promote the process of soybean salt-tolerant breeding. In the study, the membership function method was used to evaluate seven traits related to salt tolerance and comprehensive salt tolerance at the soybean seedling stage; genome-wide association analysis (GWAS) was performed in a natural population containing 200 soybean materials; and linkage analysis was performed in 112 recombinant inbred lines (RIL) population to detect quantitative trait loci (QTLs) of salt tolerance. In the GWAS, 147 SNPs were mapped, explaining 5.28-17.16% of phenotypic variation. In the linkage analysis, 10 QTLs were identified, which could explain 6.9-16.16% of phenotypic variation. And it was found that there were two co-located regions between the natural population and the RIL population, containing seven candidate genes of salt tolerance in soybean. In addition, one colocalization interval was found to contain qZJS-15-1, rs47665107, and rs4793412, all of which could explain more than 10% of phenotypic variation rates, making it suitable for molecular marker development. The physical positions of rs47665107 and rs47934112 were included in qZJS-15-1. Therefore, a KASP marker was designed and developed using Chr. 15:47907445, which was closely linked to the qZJS-15-1. This marker could accurately and clearly cluster the materials of salt-tolerant genotypes in the heterozygous population tested. The QTLs and KASP markers found in the study provide a theoretical and technical basis for accelerating the salt-tolerant breeding of soybean.
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  • 文章类型: Journal Article
    甲状腺乳头状癌(PTC),滤泡细胞衍生的最常见恶性肿瘤,通常与良好的预后有关。然而,重要的是要确定患者的侵袭性PTC和不利的结果。分子标志物如BRAFV600E突变和TERT启动子突变已被提出用于风险分层。虽然TERT启动子突变经常与侵袭性PTC相关,BRAFV600E突变与复发和死亡率增加的关联尚不清楚,目前已有争议.本研究的目的是分析差异表达基因是否可以预测PTC中的BRAFV600E突变以及TERT启动子突变。RNA测序鉴定了BRAFV600E和BRAF野生型PTC之间的大量差异表达基因。其中,AHNAK2,DCSTAMP,使用定量RT-PCR,可以在更大的队列(n=91)中证实FN1在BRAFV600E突变PTC中显著上调。此外,DCSTAMP和FN1的个体PTC表达值能够以高灵敏度和特异性预测BRAFV600E突变状态。在所有具有TERT启动子突变的PTC中和在19%的没有TERT启动子突变的PTC中检测到TERT的表达。同时具有TERT表达和TERT启动子突变的肿瘤与侵袭性临床病理特征和较短的无复发生存期特别相关。总之,探索AHNAK2,DCSTAMP,和PTC中的FN1更详细。对它们的表达模式的分析可以允许表征PTC亚型,从而使得能够进行更个性化的手术和医学治疗。
    Papillary thyroid carcinoma (PTC), the most common malignancy of follicular cell derivation, is generally associated with good prognosis. Nevertheless, it is important to identify patients with aggressive PTCs and unfavorable outcome. Molecular markers such as BRAFV600E mutation and TERT promoter mutations have been proposed for risk stratification. While TERT promoter mutations have been frequently associated with aggressive PTCs, the association of BRAFV600E mutation with increased recurrence and mortality is less clear and has been controversially discussed. The aim of the present study was to analyze whether differentially expressed genes can predict BRAFV600E mutations as well as TERT promoter mutations in PTCs. RNA sequencing identified a large number of differentially expressed genes between BRAFV600E and BRAFwildtype PTCs. Of those, AHNAK2, DCSTAMP, and FN1 could be confirmed in a larger cohort (n = 91) to be significantly upregulated in BRAFV600E mutant PTCs using quantitative RT-PCR. Moreover, individual PTC expression values of DCSTAMP and FN1 were able to predict the BRAFV600E mutation status with high sensitivity and specificity. The expression of TERT was detected in all PTCs harboring TERT promoter mutations and in 19% of PTCs without TERT promoter mutations. Tumors with both TERT expression and TERT promoter mutations were particularly associated with aggressive clinicopathological features and a shorter recurrence-free survival. Altogether, it will be interesting to explore the biological function of AHNAK2, DCSTAMP, and FN1 in PTC in more detail. The analysis of their expression patterns could allow the characterization of PTC subtypes and thus enabling a more individualized surgical and medical treatment.
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  • 文章类型: Journal Article
    在马铃薯生产中,由solani链格孢菌引起的早疫病是一种破坏性疾病。这里,通过系统地筛选由115个小的含半胱氨酸的候选Aex(交替细胞外蛋白)组成的效应蛋白库。我们鉴定了一种名为Aex59的核心效应蛋白,一种病原体相关分子模式(PAMP)分子。Aex59在真菌的子囊中独特存在,可以激活多种植物的防御反应。靶向基因破坏表明,Aex59是一种毒力因子,参与孢子发育。烟草中Aex59的感知不依赖于受体样激酶油菜素类固醇相关激酶1(BAK1)和BIR1-1抑制剂(SOBIR1),是多种模式识别受体(PRR)途径所必需的。序列分析显示,Aex59是Alta1蛋白家族的新成员,是能够帮助分类真菌的潜在分子标记。
    Early blight caused by Alternaria solani is a destructive disease in potato production. Here, through systematically screening of an effector protein pool consisting of 115 small cysteine-containing candidate Aex (Alternariaextracellular proteins) in A. solani, we identified a core effector protein named Aex59, a pathogen-associated molecular pattern (PAMP) molecule. Aex59 is uniquely present in the Ascomycota of fungi and can activate defense responses in multiple plants. Targeted gene disruption showed that Aex59 is a virulence factor and participates in spore development. Perception of Aex59 in Nicotiana benthamiana does not depend on the receptor-like kinases Brassinosteroid-associated kinase1 (BAK1) and Suppressor of BIR1-1 (SOBIR1), which are required for multiple pattern recognition receptors (PRR) pathways. Sequence analysis revealed that Aex59 is a new member of the Alt a 1 protein family and is a potential molecular marker capable of aiding in the classification of the fungi Alternaria spp.
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  • 文章类型: Journal Article
    许多研究表明,m6A在肿瘤的发展中起着重要的调节作用。HNRNPA2B1,m6ARNA甲基化阅读蛋白之一,已被证明在人类癌症中升高。
    在这项研究中,我们旨在确定HNRNPA2B1在乳腺癌中的作用。
    通过RT-qPCR和TCGA数据库研究了乳腺癌中HNRNPA2B1的表达。然后,通过CCK8检测HNRNPA2B1对癌细胞的功能,集落形成和划痕测定。此外,通过Wilcoxon符号秩检验探索BRCA中HNRNPA2B1的表达,KruskalWallis检验和逻辑回归。通过KaplanMeier和Cox回归分析考虑与HNRNPA2B1表达和存活的关联。通过基因集富集分析(GSEA)和ClusterProfilerR软件包分析HNRNPA2B1的生物学功能。
    我们发现HNRNPA2B1在乳腺癌中高表达并诱导细胞增殖和迁移。此外,我们观察到HNRNPA2B1在体内诱导肿瘤生长。此外,我们还发现HNRNPA2B1的表达与乳腺癌患者的特征和预后相关.
    我们的研究结果表明,HNRNPA2B1促进肿瘤生长,并可能作为乳腺癌新的潜在分子标志物发挥作用。
    UNASSIGNED: Numerous studies have shown that m6A plays an important regulatory role in the development of tumors. HNRNPA2B1, one of the m6A RNA methylation reading proteins, has been proven to be elevated in human cancers.
    UNASSIGNED: In this study, we aimed to identify the role of HNRNPA2B1 in breast cancer.
    UNASSIGNED: HNRNPA2B1 expression was investigated via RT-qPCR and TCGA database in breast cancer. Then, the function of HNRNPA2B1 on cancer cell was measured by CCK8 assays, colony formation and scratch assays. In addition, HNRNPA2B1 expression in BRCA was explored via the Wilcoxon signed-rank test, KruskalWallis test and logistic regression. The association with HNRNPA2B1 expression and survival were considered by KaplanMeier and Cox regression analyses. The biological function of HNRNPA2B1 was analyzed via gene set enrichment analysis (GSEA) and the cluster Profiler R software package.
    UNASSIGNED: We found that HNRNPA2B1 was highly expressed and induced cell proliferation and migration in breast cancer. Moreover, we observed HNRNPA2B1 induced tumor growth in vivo. In addition, we also found HNRNPA2B1 expression was associated with characteristics and prognosis in breast cancer patients.
    UNASSIGNED: Our findings suggested that HNRNPA2B1 promoted tumor growth and could function as a new potential molecular marker in breast cancer.
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  • 文章类型: Journal Article
    糯玉米主要在韩国种植,用于生产食品和小吃,花青素含量增加的有色玉米用于生产功能性食品和药用产品。支持关联作图分析(AMA)作为鉴定与复杂性状相关的遗传标记的优选方法。我们的研究旨在鉴定通过AMA评估的有色糯玉米近等基因系(NIL)中与两种花色苷含量和六种种皮颜色性状相关的分子标记。我们对10个有色糯玉米NIL中的285个SSR位点和两个花色苷含量和六个种皮颜色性状进行了AMA。在人口结构和集群形成的分析中,“Mibaek2ho”品种糯玉米的两个亲本系(HW3,HW9)和10个NIL明确分为两组,每组包含两个亲本近交系之一。在AMA中,在10个NILs中,62个SSR标记与2个种子花色苷含量和6个种皮颜色性状相关。所有花色苷含量和种皮颜色性状均与SSR标记相关,每个特征范围从2到12个SSR标记。12个SSR标记与两个花色苷含量(kuromanin和牡丹)性状都相关。我们目前的结果证明了SSR分析对遗传多样性检查的有效性,关系,10个有色糯玉米和“Mibaek2ho”品种糯玉米的两个亲本系的种群结构和AMA。
    Waxy maize is mainly cultivated in South Korea for the production of food and snacks, and colored maize with increased anthocyanin content is used in the production of functional foods and medicinal products. Association mapping analysis (AMA) is supported as the preferred method for identifying genetic markers associated with complex traits. Our study aimed to identify molecular markers associated with two anthocyanin content and six seed coat color traits in near-isogenic lines (NILs) of colored waxy maize assessed through AMA. We performed AMA for 285 SSR loci and two anthocyanin content and six seed coat color traits in 10 NILs of colored waxy maize. In the analysis of population structure and cluster formation, the two parental lines (HW3, HW9) of \"Mibaek 2ho\" variety waxy maize and the 10 NILs were clearly divided into two groups, with each group containing one of the two parental inbred lines. In the AMA, 62 SSR markers were associated with two seed anthocyanin content and six seed coat color traits in the 10 NILs. All the anthocyanin content and seed coat color traits were associated with SSR markers, ranging from 2 to 12 SSR markers per characteristic. The 12 SSR markers were together associated with both of the two anthocyanin content (kuromanin and peonidin) traits. Our current results demonstrate the effectiveness of SSR analysis for the examination of genetic diversity, relationships, and population structure and AMA in 10 NILs of colored waxy maize and the two parental lines of the \"Mibaek 2ho\" variety waxy maize.
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  • 文章类型: Editorial
    暂无摘要。
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  • 文章类型: Journal Article
    土壤细菌对不溶性磷化合物的溶解具有重要意义,因为它使植物可以使用磷。有机酸的产生是溶解不溶性无机磷化合物的主要微生物机理。在革兰氏阴性细菌中,葡萄糖酸是由葡萄糖脱氢酶-吡咯并喹啉奎宁全酶GDH-PQQ的活性合成的。标记基因的使用是评估引入细菌的持久性并允许跟踪生物和非生物因素对土壤中这些有益微生物的影响的非常有用的工具。在先前的研究中,我们在很大一部分不可培养和可培养的天然土壤细菌中检测到pqqE基因的存在。这项研究的目的是分析pqqE基因序列的系统发育及其在纯细菌和混合细菌培养物以及根际土壤样品中研究解磷细菌的潜力。为此,通过PCR确定属于几种细菌的磷酸盐溶解细菌基因组中pqqE基因的存在。此外,该基因是从与接种或未接种磷酸盐溶解细菌的花生植物相关的混合细菌培养物和根际土壤中进行分析的。为此,从几个细菌属设计的简并引物和假单胞菌属的特异性引物。,在这项研究中设计的,被使用。使用两种不同的DNA提取技术从简单或混合的细菌培养物以及根际土壤样品中获得DNA模板。结果表明,在所有革兰氏阴性磷酸盐溶解细菌的基因组中均发现了pqqE基因扩增产物。有可能在从这些细菌与其他微生物相互作用的混合培养物中获得的DNA中以及从接种或未接种这些细菌的根际土壤样品中获得的DNA中检测到该基因。系统发育分析表明,pqqE基因是相关属中的保守基因。总之,pqqE基因可能是研究磷酸盐溶解细菌种群的潜在标记。
    Insoluble phosphorous compounds solubilization by soil bacteria is of great relevance since it puts available the phosphorus to be used by plants. The production of organic acids is the main microbiological mechanism by which insoluble inorganic phosphorus compounds are solubilized. In Gram negative bacteria, gluconic acid is synthesized by the activity of the holoenzyme glucose dehydrogenase-pyrroloquinoline quinine named GDH-PQQ. The use of marker genes is a very useful tool to evaluate the persistence of the introduced bacteria and allow to follow-up the effect of biotic and abiotic factors on these beneficial microorganisms in the soil. In previous studies we detected the presence of the pqqE gene in a great percentage of both non-culturable and culturable native soil bacteria. The objective of this study was to analyze the phylogeny of the sequence of pqqE gene and its potential for the study of phosphate solubilizing bacteria from pure and mixed bacterial cultures and rhizospheric soil samples. For this, the presence of the pqqE gene in the genome of phosphate solubilizing bacteria that belong to several bacteria was determined by PCR. Also, this gene was analyzed from mixed bacterial cultures and rhizospheric soil associated to peanut plants inoculated or not with phosphate solubilizing bacteria. For this, degenerate primers designed from several bacterial genera and specific primers for the genus Pseudomonas spp., designed in this study, were used. DNA template used from simple or mixed bacterial cultures and from rhizospheric soil samples was obtained using two different DNA extraction techniques. Results indicated that pqqE gene amplification product was found in the genome of all Gram negative phosphate solubilizing bacteria analyzed. It was possible to detect this gene in the DNA obtained from mixed cultures where these bacteria grew in interaction with other microorganisms and in that obtained from rhizospheric soil samples inoculated or not with these bacteria. The phylogenetic analysis indicated that pqqE gene is a conserved gene within related genera. In conclusion, pqqE gene could be a potential marker for the study of phosphate solubilizing bacterial populations.
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  • 文章类型: Journal Article
    籽粒垩白直接影响稻米的商业价值。到目前为止,已经在突变体中发现了与白垩相关的基因,但是尚未确定这些基因是否可以通过育种来改善水稻品质。因此,在水稻种质中发现更多的数量性状位点(QTLs)或与垩白相关的基因是必要的。这项研究需要通过将120万个单核苷酸多态性(SNP)与173个水稻品种的表型数据相结合,对胚乳白垩度(DEC)和白垩的谷物百分比(PGWC)进行全基因组关联研究。确定了DEC的13个QTL和PGWC的9个QTL,其中四个同时检测到DEC和PGWC;此外,qDEC11/qPGWC11被鉴定为主要的QTL。通过结合连锁不平衡分析和SNP信息,LOC_Os11g10170被鉴定为DEC的候选基因。LOC_Os11g10170的单倍型之间存在显着差异,并且观察到LOC_Os11g10170的Hap1使DEC降低了6.19%。qRT-PCR结果表明,在开花后第21-42天,高DEC值的种质的基因表达水平显着高于低DEC值的种质。最长为28天。这些结果为水稻白垩相关性状的遗传改良提供了分子标记和种质资源。
    Grain chalkiness directly affects the commercial value of rice. Genes related to chalkiness reported thus far have been discovered in mutants, but it has not been identified whether these genes can be used to improve rice quality by breeding. Therefore, discovering more quantitative trait loci (QTLs) or genes related to chalkiness in the rice germplasm is necessary. This study entails a genome-wide association study on the degree of endosperm chalkiness (DEC) and percentage of grains with chalkiness (PGWC) by combining 1.2 million single-nucleotide polymorphisms (SNPs) with the phenotypic data of 173 rice accessions. Thirteen QTLs for DEC and nine for PGWC were identified, of which four were detected simultaneously for both DEC and PGWC; further, qDEC11/qPGWC11 was identified as the major QTL. By combining linkage disequilibrium analysis and SNP information, LOC_Os11g10170 was identified as the candidate gene for DEC. There were significant differences among the haplotypes of LOC_Os11g10170, and the Hap 1 of LOC_Os11g10170 was observed to reduce the DEC by 6.19%. The qRT-PCR results showed that the gene expression levels in accessions with high DEC values were significantly higher than those in accessions with low DEC values during days 21-42 after flowering, with a maximum at 28 days. These results provide molecular markers and germplasm resources for genetic improvement of the chalkiness-related traits in rice.
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  • 文章类型: Journal Article
    尽管对氨基水杨酸(PAS)已被用于治疗结核病数十年,结核分枝杆菌对这种药物的耐药机制(M.结核病)尚未对临床分离株进行彻底调查。以前,我们发现,Rv2172c亚甲基四氢叶酸还原酶(MTHFR)活性降低导致结核分枝杆菌抗叶酸药物敏感性增加.在这项研究中,我们收集了173株PAS耐药和803株PAS敏感临床分离株的基因组测序数据,并分析了这976株分离株中的rv2172c突变.结果表明,在一定比例(6.36%)的PAS抗性分离株中,可以在rv2172c上鉴定出两个突变(T120P和M172V)。AlphaFold2预测结果表明,T120P或M172V突变可能通过影响烟酰胺腺嘌呤二核苷酸(NADH)结合来影响Rv2172c的酶活性,随后的生化分析证实了这一点,证明残基Thr120和Met172对NADH结合和Rv2172c的酶活性的作用。此外,在结核分枝杆菌中确定了rv2172cT120P或M172V突变对甲硫氨酸产生和PAS抗性的影响.结果表明,T120P和M172V突变均导致细胞内蛋氨酸浓度增加和高水平的PAS抗性。总之,我们在结核分枝杆菌临床分离株中发现了新的分子标记和PAS耐药的新机制,并拓宽了对结核分枝杆菌中Rv2172c的NADH依赖性MTHFR催化机制的理解,这将有助于PAS耐药性的分子诊断以及靶向Rv2172c的新药的开发。
    Although para-aminosalicylic acid (PAS) has been used to treat tuberculosis for decades, mechanisms of resistance to this drug in Mycobacterium tuberculosis (M. tuberculosis) clinical isolates have not been thoroughly investigated. Previously, we found that decreased methylenetetrahydrofolate reductase (MTHFR) activity of Rv2172c led to increased sensitivity to antifolates in M. tuberculosis. In this study, we collected the genome-sequencing data of 173 PAS-resistant and 803 PAS-sensitive clinical isolates and analyzed rv2172c mutations in those 976 isolates. The results showed that two mutations (T120P and M172V) on rv2172c could be identified in a certain proportion (6.36%) of PAS-resistant isolates. The results of AlphaFold2 prediction indicated that the T120P or M172V mutation might affect the enzymatic activity of Rv2172c by influencing nicotinamide adenine dinucleotide (NADH) binding, and this was verified by subsequent biochemical analysis, demonstrating the role of residues Thr120 and Met172 on NADH binding and enzymatic activity of Rv2172c. In addition, the effect of rv2172c T120P or M172V mutation on methionine production and PAS resistance was determined in M. tuberculosis. The results showed that both T120P and M172V mutations caused increased intracellular methionine concentrations and high level PAS resistance. In summary, we discovered new molecular markers and also a novel mechanism of PAS resistance in M. tuberculosis clinical isolates and broadened the understanding of the NADH-dependent MTHFR catalytic mechanism of Rv2172c in M. tuberculosis, which will facilitate the molecular diagnosis of PAS resistance and also the development of new drugs targeting Rv2172c.
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  • 文章类型: Journal Article
    PhysalisL.属植物已被广泛用于传统和本土的中药治疗各种疾病,包括皮炎,疟疾,哮喘,肝炎,和肝脏疾病。本综述旨在对酸浆属进行全面和最新的调查,一种新型作物,了解植物多样性和果实发育。几种叶绿体DNA-,核核糖体DNA-,和基于基因组DNA的标记,比如psbA-trnH,内转录间隔区(ITS),简单序列重复(SSR),随机扩增微卫星(RAMS),序列特征扩增区(SCAR),和单核苷酸多态性(SNP),被开发用于分子鉴定,遗传多样性,和酸浆物种的系统发育研究。大量的功能基因参与了花萼膨胀综合征的发展(AP2-L,MPF2、MPF3和MAGO),器官生长(AG1,AG2,POS1和CNR1),和活性成分代谢(24ISO,DHRT,P450-CPL,SR,鉴定出DUF538,TAS14和3β-HSB)有助于新型酸浆品种的育种。各种组学研究揭示并在功能上确定了一系列生殖器官发育相关因素,环境应激反应基因,和活性成分生物合成相关酶。佛罗里达酸浆Rydb的染色体水平基因组。,酸浆(Waterf。)M.马丁内斯,和酸浆最近已经出版,为酸浆作物的基因组编辑提供了宝贵的资源。我们的综述总结了遗传多样性的最新进展,分子鉴定,系统发育学,功能基因,以及组学在酸浆属中的应用,并加速了这种传统草药的有效利用。
    The plants of the genus Physalis L. have been extensively utilized in traditional and indigenous Chinese medicinal practices for treating a variety of ailments, including dermatitis, malaria, asthma, hepatitis, and liver disorders. The present review aims to achieve a comprehensive and up-to-date investigation of the genus Physalis, a new model crop, to understand plant diversity and fruit development. Several chloroplast DNA-, nuclear ribosomal DNA-, and genomic DNA-based markers, such as psbA-trnH, internal-transcribed spacer (ITS), simple sequence repeat (SSR), random amplified microsatellites (RAMS), sequence-characterized amplified region (SCAR), and single nucleotide polymorphism (SNP), were developed for molecular identification, genetic diversity, and phylogenetic studies of Physalis species. A large number of functional genes involved in inflated calyx syndrome development (AP2-L, MPF2, MPF3, and MAGO), organ growth (AG1, AG2, POS1, and CNR1), and active ingredient metabolism (24ISO, DHCRT, P450-CPL, SR, DUF538, TAS14, and 3β-HSB) were identified contributing to the breeding of novel Physalis varieties. Various omic studies revealed and functionally identified a series of reproductive organ development-related factors, environmental stress-responsive genes, and active component biosynthesis-related enzymes. The chromosome-level genomes of Physalis floridana Rydb., Physalis grisea (Waterf.) M. Martínez, and Physalis pruinosa L. have been recently published providing a valuable resource for genome editing in Physalis crops. Our review summarizes the recent progress in genetic diversity, molecular identification, phylogenetics, functional genes, and the application of omics in the genus Physalis and accelerates efficient utilization of this traditional herb.
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