神奇“魔法条”之新冠病毒的快速检测

胶体金作为商业化的即时检验POCT(point-of-care test)免疫层析技术中最广泛的标记物之一,具有显著的颜色特征,后期无需可视化改造,且形态稳定、环境友好。本文试纸条制作过程简单,不需要大型贵重仪器。采用柠檬酸三钠还原氯金酸制备胶体金,将胶体金标记在新冠病毒的核衣壳N蛋白抗体上,通过在免疫层析试纸条可视区形成类似三明治结构的抗体-抗原-抗体夹心复合物,并以此复合物的红色检测线作为新冠病毒的定性分析依据。该检测方法裸眼可测、操作简单、且快速(10 min)、灵敏(检测限为0.5 ng∙mL^(−1))、低成本(每条不超过5元)。实验过程可以引导学生体验生命体系中抗原抗体相互作用引发的特异性生命化学反应,激发学生对化学、生物、生命科学和纳米材料等学科相互融合的可视化快检技术的浓厚研究兴趣。

The transcription factor IbNAC29 positively regulates the carotenoid accumulation in sweet potato

Carotenoid is a tetraterpene pigment beneficial for human health.Although the carotenoid biosynthesis pathway has been extensively studied in plants,relatively little is known about their regulation in sweet potato.Previously,we conducted the transcriptome database of differentially expressed genes between the sweet potato(Ipomoea batatas)cultivar‘Weiduoli’and its high-carotenoid mutant‘HVB-3’.In this study,we selected one of these candidate genes,IbNAC29,for subsequent analyses.IbNAC29 belongs to the plant-specific NAC(NAM,ATAF1/2,and CUC2)transcription factor family.Relative IbNAC29 mRNA level in the HVB-3 storage roots was∼1.71-fold higher than Weiduoli.Additional experiments showed that the contents ofα-carotene,lutein,β-carotene,zeaxanthin,and capsanthin are obviously increased in the storage roots of transgenic sweet potato plants overexpressing IbNAC29.Moreover,the levels of carotenoid biosynthesis genes in transgenic plants were also up-regulated.Nevertheless,yeast one-hybrid assays indicated that IbNAC29 could not directly bind to the promoters of these carotenoid biosynthesis genes.Furthermore,the level of IbSGR1 was down-regulated,whose homologous genes in tomato can negatively regulate carotene accumulation.Yeast three-hybrid analysis revealed that the IbNAC29-IbMYB1R1-IbAITR5 could form a regulatory module.Yeast one-hybrid,electrophoretic mobility shift assay,quantitative PCR analysis of chromatin immunoprecipitation and dual-luciferase reporter assay showed that IbAITR5 directly binds to and inhibits the promoter activity of IbSGR1,up-regulating carotenoid biosynthesis gene IbPSY.Taken together,IbNAC29 is a potential candidate gene for the genetic improvement of nutritive value in sweet potato.

Construction of a high-density SSR genetic linkage map and identification of QTL for storage-root yield and dry-matter content in sweetpotato

Sweetpotato(Ipomoea batatas(L.)Lam.)is a widely grown food crop especially in developing countries.Increasing storage-root yield and dry-matter content has been the main breeding objective of the crop,and DNA marker-assisted breeding is needed for this purpose.In this study,using a mapping population of 500 F1 individuals from a cross between Xushu 18(female)and Xu 781(male),we constructed a highdensity genetic linkage map of sweetpotato using 601 simple-sequence repeat(SSR)primer pairs.The Xushu 18 map contained 90 linkage groups with 5547 SSR markers and spanned 18,263.5 cM,and the Xu 781 map contained 90 linkage groups with 4599 SSR markers and spanned 18,043.7 cM,representing the highest genome coverage yet reported for sweetpotato.We identified 33 QTL for storage-root yield and 16 QTL for dry-matter content,explaining respectively 6.5%–47.5%and 3.2%–18.9%of variation.These results provide a foundation for fine-mapping and cloning of QTL and for marker-assisted breeding in sweetpotato.

Regulation of soybean stem growth habit:A ten-year progress report

Stem growth habit dictates plant architecture and influences flowering and podding(seed setting),making it an essential morphological and breeding agronomic trait of soybean(Glycine max).Stem growth habit in soybean is affected by photoperiod and environment and is determined by genetic variation at major genes.Classical genetic analysis identified two critical loci,designated Determinacy 1(Dt1)and Determinacy 2(Dt2).Dt1 is an ortholog of Arabidopsis thaliana TERMINAL FLOWER1(TFL1)and specifies an indeterminate stem growth habit,whereas Dt2 specifies a semi-determinate growth habit.MADS-box proteins,including Dt2,SUPPRESSOR OF OVEREXPRESSION OF CO1(GmSOC1)and MADS-box genes downregulated by E1(GmMDE),repress Dt1 expression.Photoreceptors encoded by the E3 and E4 loci regulate the expression of soybean FLOWERING LOCUS T(GmFT)orthologs via circadian clock genes and E1,and GmFTs compete with Dt1 to regulate stem growth habit.Study of the molecular mechanism underlying the regulation of stem growth habit in soybean has focused on the repression of Dt1 expression.Here we provide an overview of progress made in elucidating the genetic and molecular bases of stem growth habit in soybean,with emphasis on the molecular components responsible for integrating photoperiodic flowering and stem growth habit.

Development of a high-density SSR genetic linkage map in sweet potato

Simple sequence repeat(SSR) markers have been proved to be a very powerful tool for quantitative trait locus(QTL) mapping, marker-assisted selection and comparative genomics research in many crop species. However, a high-density SSR genetic linkage map is still lacking because there are only a few SSR markers available in sweet potato. In this study, a total of 2545 simple sequence repeat(SSR) primer pairs, including 1215 genomic SSR(gSSR) primer pairs and 1330 BES-SSR(bSSR) primer pairs designed from the genome sequence and BAC-end sequence of sweet potato, respectively, were screened with sweet potato cultivars Luoxushu 8 and Zhengshu 20 and their randomly sampled two F1 individuals and 571 of them generated polymorphic bands. The selected 571 polymorphic SSR primer pairs and 35 EST-based SSR(eSSR) primer pairs developed at our laboratory were used to genotype 240 F1 individuals derived from a cross between Luoxushu 8 and Zhengshu 20. A double pseudo-test-cross strategy was applied for linkage analysis. The Luoxushu 8 map included 90 linkage groups with 5057 SSR markers and covered 13,299.9 cM with a marker density of 2.6 cM, and the Zhengshu 20 map contained 90 linkage groups with 3009 SSR markers and covered 11,122.9 cM with a marker density of 3.7 cM. Fifteen homologous groups were identified in both parent maps. These are the first SSR linkage maps consisting of the complete 90 linkage groups and 15 homologous groups, which are consistent with the autohexaploid nature of sweetpotato, and are also the linkage maps with the highest SSR marker density reported to date.These results provide a basis for QTL mapping, marker-assisted breeding and comparative genomics research of sweet potato.

A sucrose non-fermenting-1-related protein kinase-1 gene, IbSnRK1, confers salt, drought and cold tolerance in sweet potato

Sucrose non-fermenting-1-related protein kinase-1(SnRK1)regulates carbon and nitrogen metabolism in plants.However,its roles and their underlying mechanisms in tolerance to abiotic stresses are little known.The present study indicated that the IbSnRK1 gene was strongly induced by Na Cl,polyethylene glycol(PEG)6000,hydrogen peroxide(H2O2),cold(4°C),and abscisic acid(ABA).Its overexpression significantly increased salt,drought,and cold tolerance in transgenic sweet potato plants.ABA,proline,and K+contents were significantly increased,whereas malondialdehyde(MDA),Na+and H2O2 contents and O2-production rate were significantly decreased in the transgenic plants under salt,drought,and cold stresses.Overexpression of the gene up-regulated genes involved in ABA biosynthesis,stress response,and stomatal closure;increased enzyme activities in the reactive oxygen species(ROS)scavenging system;and controlled stomatal closure under salt,drought,and cold stresses.These results show that the IbSnRK1 gene confers salt,drought,and cold tolerance in sweet potato by activating the ROS scavenging system and controlling stomatal closure via the ABA signaling pathway.

A critical role of PvFtsH2 in the degradation of photodamaged D1 protein in common bean

Light is required for initiating chloroplast biogenesis and photosynthesis;however,the photosystem II reaction center(PSII RC)can be photodamaged.In this study,we characterized pvsl1,a seedling-lethal mutant of Phaseolus vulgaris.This mutant showed lethality when exposed to sunlight irradiation and a yellow-green leaf phenotype when grown in a growth chamber under low-light conditions.We developed 124 insertion/deletion(INDEL)markers based on resequencing data of Dalong1 and PI60234,two local Chinese common bean cultivars,for genetic mapping.We identified Phvul.002G190900,which encodes the PvFtsH2 protein,as the candidate gene for this pvsl1 mutation through fine-mapping and functional analysis.A single-base deletion occurred in the coding region of Phvul.002G190900 in the pvsl1 mutant,resulting in a frameshift mutation and a truncated protein lacking the Zn^(2+)metalloprotease domain.Suppressed expression of Phvul.002G190900 at the transcriptional level was detected,while no change in the subcellular localization signal was observed.The seedlings of pvsl1 exhibited hypersensitivity to photoinhibition stress.In the pvsl1 mutant,abnormal accumulation of the D1 protein indicated a failure to rapidly degrade damaged D1 protein in the PSII RC.The results of this study demonstrated that PvFtsH2 is critically required for survival and maintaining photosynthetic activity by degrading photodamaged PSII RC D1 protein in common bean.

IbNIEL-mediated degradation of IbNAC087 regulates jasmonic acid-dependent salt and drought tolerance in sweet potato

Sweet potato(Ipomoea batatas[L.]Lam.)is a crucial staple and bioenergy crop.Its abiotic stress tolerance holds significant importance in fully utilizing marginal lands.Transcriptional processes regulate abiotic stress responses,yet the molecular regulatory mechanisms in sweet potato remain unclear.In this study,a NAC(NAM,ATAF1/2,and CUC2)transcription factor,IbNAC087,was identified,which is commonly upregulated in salt-and drought-tolerant germplasms.Overexpression of IbNAC087 increased salt and drought tolerance by increasing jasmonic acid(JA)accumulation and activating reactive oxygen species(ROS)scavenging,whereas silencing this gene resulted in opposite phenotypes.JA-rich IbNAC087-OE(overexpression)plants exhibited more stomatal closure than wild-type(WT)and IbNAC087-Ri plants under NaCl,polyethylene glycol,and methyl jasmonate treatments.IbNAC087 functions as a nuclear transcriptional activator and directly activates the expression of the key JA biosynthesisrelated genes lipoxygenase(IbLOX)and allene oxide synthase(IbAOS).Moreover,IbNAC087 physically interacted with a RING-type E3 ubiquitin ligase NAC087-INTERACTING E3 LIGASE(IbNIEL),negatively regulating salt and drought tolerance in sweet potato.IbNIEL ubiquitinated IbNAC087 to promote 26S proteasome degradation,which weakened its activation on IbLOX and IbAOS.The findings provide insights into the mechanism underlying the IbNIEL-IbNAC087 module regulation of JA-dependent salt and drought response in sweet potato and provide candidate genes for improving abiotic stress tolerance in crops.

A novel aldo-keto reductase gene, IbAKR, from sweet potato confers higher tolerance to cadmium stress in tobacco

High concentrations of Cd can inhibit growth and reduce the activity of the photosynthetic apparatus in plants. In several plant species, aldo-keto reductases(AKRs) have been shown to enhance tolerance to various abiotic stresses by scavenging cytotoxic aldehydes;however, few AKRs have been reported to enhance Cd stress tolerance. In this study, the gene Ib AKR was isolated from sweet potato. The relative expression levels of Ib AKR increased significantly(approximately 3-fold) after exposure to 200 mmol$L–1 Cd Cl2 or 10 mmol$L–1 H2 O2. A subcellular localization assay showed that Ib AKR is predominantly located in the nucleus and cytoplasm.Ib AKR-overexpressing tobacco plants showed higher tolerance to Cd stress than wild-type(WT). Transgenic lines showed a significant ability to scavenge malondialdehyde(MDA) and methylglyoxal(MG). In addition,proline content and superoxide dismutase activity were significantly higher and H2 O2 levels were significantly lower in the transgenic plants than in the WT. Quantitative real-time PCR analysis showed that the reactive oxygen species(ROS) scavenging genes encoding guaiacol peroxidase(GPX), ascorbate peroxidase(APX), monodehydroascorbate reductase(MDHAR) and peroxidase(POD) were significantly upregulated in transgenic plants compared to WT under Cd stress. These findings suggest that overexpressing Ib AKR enhances tolerance to Cd stress via the scavenging of cytotoxic aldehydes and the activation of the ROS scavenging system.

Comparative transcriptome analysis of purple-fleshed sweet potato provides insights into the molecular mechanism of anthocyanin biosynthesis

Sweet potato, Ipomoea batatas, is a globally important food crop. The purple-fleshed sweet potato, rich in anthocyanins, has great potential for both nutritional and pharmaceutical uses. In this study, we characterized the root transcriptomes of the purple-fleshed sweet potato cv.Jingshu 6 and its mutant JS6-5 with high anthocyanin content by high-throughput RNA sequencing. A total of22873364 and 27955097 high quality reads were obtained from Jingshu 6 and JS6-5, respectively, and assembled into35592 unigenes. In all, we obtained 1566 differentially expressed genes(DEGs). Among them, 994 were upregulated and 572 were downregulated in JS6-5 compared to the expression in Jingshu 6. A total of 1436 DEGs were annotated, in which 847 DEGs had gene ontology(GO) terms and 329 DEGs were assigned to 84 Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways. Most importantly, 23 differentially expressed genes and 24 transcription factors were identified as candidate genes involved in anthocyanin biosynthesis. In addition, 2349 SSRs were detected. This study not only provides the candidate genes but also provides insights into the molecular mechanism of anthocyanin biosynthesis in sweet potato.

QNE1 is a key flowering regulator determining the length of the vegetative period in soybean cultivars

The soybean E1 gene is a major regulator that plays an important role in flowering time and maturity.However,it remains unclear how cultivars carrying the dominant E1 allele adapt to the higher latitudinal areas of northern China.We mapped the novel quantitative trait locus QNE1(QTL near E1) for flowering time to the region proximal to E1 on chromosome 6 in two mapping populations.Positional cloning revealed Glyma.06G204300,encoding a TCP-type transcription factor,as a strong candidate gene for QNE1.Association analysis further confirmed that functional single nucleotide polymorphisms(SNPs) at nucleotides 686 and 1,063 in the coding region of Glyma.06G204300 were significantly associated with flowering time.The protein encoded by the candidate gene is localized primarily to the nucleus.Furthermore,soybean and Brassica napus plants overexpressing Glyma.06G204300 exhibited early flowering.We conclude that despite their similar effects on flowering time,QNE1 and E4 may control flowering time through different regulatory mechanisms,based on expression studies and weighted gene co-expression network analysis of flowering time-related genes.Deciphering the molecular basis of QNE1 control of flowering time enriches our knowledge of flowering gene networks in soybean and will facilitate breeding soybean cultivars with broader latitudinal adaptation.