CLC number:
On-line Access: 2021-06-11
Received: 2020-09-10
Revision Accepted: 2020-11-10
Crosschecked: 0000-00-00
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Citations: Bibtex RefMan EndNote GB/T7714
https://orcid.org/0000-0002-1612-5620
Yun WU, Minyi SUN, Shiqi LI, Ruihan MIN, Cong GAO, Qundan LYU, Ziming REN, Yiping XIA. Molecular cloning, characterization and expression analysis of three key starch synthesis-related genes from the bulb of a rare lily germplasm, Lilium brownii var. giganteum[J]. Journal of Zhejiang University Science B,in press.Frontiers of Information Technology & Electronic Engineering,in press.https://doi.org/10.1631/jzus.B2000545 @article{title="Molecular cloning, characterization and expression analysis of three key starch synthesis-related genes from the bulb of a rare lily germplasm, Lilium brownii var. giganteum", %0 Journal Article TY - JOUR
特异性种质巨球百合鳞茎淀粉合成关键酶基因的克隆、鉴定及表达分析创新点:本研究首次从重要野生百合资源中系统克隆淀粉合成关键酶基因,并分析其时空表达特性,获得了AGPS基因候选单核苷酸多态性(SNP)位点。 方法:采用蒽酮法测定非结构性碳水化合物含量并用高碘酸-希夫反应染色法及投射电镜观察淀粉的显微及超微结构;采用cDNA末端快速扩增(RACE)技术克隆获得巨球百合淀粉合成相关基因,并应用BLAST、ExPASy、TMHMM、SWISS-MODE、Cell-PLoc、SMART、MEGA及KaKs_Calculator等生物信息学软件对其基因序列进行分析;采用实时荧光定量PCR分析基因的时空表达差异。 结论:本研究证实巨球百合及原生种野百合在淀粉含量及特性方面存在差异,并用RACE技术克隆获得了LbgAGPS、LbgGBSS及LbgSSIII基因。三种基因均在鳞茎中表达较高,且随着鳞茎发育整体呈现上升趋势,与鳞茎的形成存在较强的相关性。AGPS基因上存在丰富的SNP位点,且主要为纯化选择。 关键词组: Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article
Reference[1]AbtMR, PfisterB, SharmaM, et al., 2020. STARCH SYNTHASE 5, a noncanonical starch synthase-like protein, promotes starch granule initiation in Arabidopsis. Plant Cell, 32(8):2543-2565. [2]ArnoldK, BordoliL, KoppP, et al., 2006. The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics, 22(2):195-201. [3]AskariN, VisserRGF, de KlerkGJ, 2018. Growth of lily bulblets in vitro, a review. Int J Hortic Sci Technol, 5(2):133-143. [4]BahajiA, LiJ, Sánchez-LópezÁM, et al., 2014. Starch biosynthesis, its regulation and biotechnological approaches to improve crop yields. Biotechnol Adv, 32(1):87-106. [5]BakhshaieM, KhosraviS, AzadiP, et al., 2016. Biotechnological advances in Lilium. Plant Cell Rep, 35(9):1799-1826. [6]BallicoraMA, LaughlinMJ, FuY, et al., 1995. Adenosine 5'-diphosphate-glucose pyrophosphorylase from potato tuber (significance of the N terminus of the small subunit for catalytic properties and heat stability). Plant Physiol, 109(1):245-251. [7]BecklesDM, CraigJ, SmithAM, 2001. ADP-glucose pyrophosphorylase is located in the plastid in developing tomato fruit. Plant Physiol, 126(1):261-266. [8]BejarCM, BallicoraMA, IglesiasAA, et al., 2006. ADPglucose pyrophosphorylase’s N-terminus: structural role in allosteric regulation. Biochem Biophys Res Commun, 343(1):216-221. [9]ChengN, ZengXF, ZhengXF, et al., 2015. Cloning and characterization of the genes encoding the small and large subunit of the ADP-glucose pyrophosphorylase in lotus (Nelumbo nucifera Gaertn). Acta Physiolo Plant, 37(1):1734. [10]ChouKC, ShenHB, 2010. Cell-PLoc 2.0: an improved package of web-servers for predicting subcellular localization of proteins in various organisms. Nat Sci, 2(10):1090-1103. [11]de KlerkGJ, 2012. Micropropagation of bulbous crops: technology and present state. Floricult Ornam Biotechnol, 6:1-8. [12]DeléageG, 2017. ALIGNSEC: viewing protein secondary structure predictions within large multiple sequence alignments. Bioinformatics, 33(24):3991-3992. [13]DoleželJ, GreilhuberJ, SudaJ, 2007. Estimation of nuclear DNA content in plants using flow cytometry. Nat Protoc, 2(9):2233-2244. [14]DuF, JiangJ, JiaHM, et al., 2015. Selection of generally applicable SSR markers for evaluation of genetic diversity and identity in Lilium. Biochem Syst Ecol, 61:278-285. [15]DuHH, YangT, MaCY, et al., 2012. Effects of RNAi silencing of SSIII gene on phosphorus content and characteristics of starch in potato tubers. J Integr Agric, 11(12):1985-1992. [16]DuYP, BiY, ZhangMF, et al., 2017. Genome size diversity in Lilium (Liliaceae) is correlated with karyotype and environmental traits. Front Plant Sci, 8:1303. [17]GanalMW, AltmannT, RöderMS, 2009. SNP identification in crop plants. Curr Opin Plant Bio, 12(2):211-217. [18]GaoMP, ZhangSW, LuoC, et al., 2018. Transcriptome analysis of starch and sucrose metabolism across bulb development in Sagittaria sagittifolia. Gene, 649:99-112. [19]GorenA, AshlockD, TetlowIJ, 2018. Starch formation inside plastids of higher plants. Protoplasma, 255(6):1855-1876. [20]GuanR, ZhaoYP, ZhangH, et al., 2016. Draft genome of the living fossil Ginkgo biloba. Gigascience, 5(1):49. [21]HuangJH, ZhouRR, HeD, et al., 2020. Rapid identification of Lilium species and polysaccharide contents based on near infrared spectroscopy and weighted partial least square method. Int J Biol Macromol, 154:182-187. [22]HuangYF, YangMX, ZhangH, et al., 2009. Genetic diversity and genetic structure analysis of the natural populations of Lilium brownii from Guangdong, China. Biochem Genet, 47(7-8):503-510. [23]KamenetskyR, OkuboH, 2012. Ornamental Geophytes: from Basic Science to Sustainable Production. CRC Press, Boca Raton, Florida, USA, p.1-16. [24]KroghA, LarssonB, von HeijneG, et al., 2001. Predicting transmembrane protein topology with a hidden markov model: application to complete genomes. J Mol Biol, 305(3):567-580. [25]KumarS, StecherG, TamuraK, 2016. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol, 33(7):1870-1874. [26]LeeGJ, SubburajS, KumariS, et al., 2016. Identification of single nucleotide polymorphism (SNP) markers associated with botrytis resistance in lily (Lilium spp.). Hortscience, 51:S320-S321. [27]LetunicI, BorkP, 2018. 20 years of the SMART protein domain annotation resource. Nucleic Acids Res, 46(D1):D493-D496. [28]LetunicI, BorkP, 2019. Interactive Tree Of Life (iTOL) v4: recent updates and new developments. Nucleic Acids Res, 47(W1):W256-W259. [29]LiGY, ChenZH, YanFB, 2007. Lilium brownin var. giganteum: a new variety of Lilium from Wenling, Zhejiang. J Zhejiang Forestry Coll, 24(6):767-768 (in Chinese). [30]LiXY, WangCX, ChengJY, et al., 2014. Transcriptome analysis of carbohydrate metabolism during bulblet formation and development in Lilium davidii var. unicolor. BMC Plant Biol, 14:358. [31]LinebargerCRL, BoehleinSK, SewellAK, et al., 2005. Heat stability of maize endosperm ADP-glucose pyrophosphorylase is enhanced by insertion of a cysteine in the N terminus of the small subunit. Plant Physiol, 139(4):1625-1634. [32]LivakKJ, SchmittgenTD, 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25(4):402-408. [33]LloydJR, KossmannJ, 2019. Starch trek: the search for yield. Front Plant Sci, 9:1930. [34]LombardV, RamuluHG, DrulaE, et al., 2014. The carbohydrate-active enzymes database (CAZy) in 2013. Nucleic Acids Res, 42(D1):D490-D495. [35]MaPP, YuanY, ShenQC, et al., 2019. Evolution and expression analysis of starch synthase gene families in Saccharum spontaneum. Trop Plant Biol, 12(3):158-173. [36]MammadovJ, AggarwalR, BuyyarapuR, et al., 2012. SNP markers and their impact on plant breeding. Int J Plant Genomics, 2012:728398. [37]MatsuoT, MizunoT, 1974. Changes in the amounts of two kinds of reserve glucose-containing polysaccharides during germination of the Easter lity bulb. Plant Cell Physiol, 15(3):555-558. [38]MiaoHX, SunPG, LiuWX, et al., 2014. Identification of genes encoding granule-bound starch synthase involved in amylose metabolism in banana fruit. PLoS ONE, 9(2):e88077. [39]MishraBP, KumarR, MohanA, et al., 2017. Conservation and divergence of Starch Synthase III genes of monocots and dicots. PLoS ONE, 12(12):e0189303. [40]Moreno-PachónN, 2017. Mechanisms of Vegetative Propagation in Bulbs: A Molecular Approach. PhD Thesis, Wageningen University, Wageningen, the Netherlands. [41]MurashigeT, SkoogF, 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant, 15(3):473-497. [42]Oteo-GarcíaG, OteoJA, 2019. Evolutionary distances corrected for purifying selection and ancestral polymorphisms. J Theor Biol, 483:110004. [43]PodwyszyńskaM, 2012. The mechanisms of in vitro storage organ formation in ornamental geophytes. Floricult Ornam Biotechnol, 6(1):9-23. [44]QuJZ, XuST, ZhangZQ, et al., 2018. Evolutionary, structural and expression analysis of core genes involved in starch synthesis. Sci Rep, 8:12736. [45]SeoSG, BeaSH, JunBK, et al., 2015. Overexpression of ADP-glucose pyrophosphorylase (IbAGPaseS) affects expression of carbohydrate regulated genes in sweet potato [Ipomoea batatas (L.) Lam. cv. Yulmi]. Genes Genom, 37(7):595-605. [46]ShahinA, van KaauwenM, EsselinkD, et al., 2012. Generation and analysis of expressed sequence tags in the extreme large genomes Lilium and Tulipa. BMC Genomics, 13:640. [47]SmithAM, ZeemanSC, 2020. Starch: a flexible, adaptable carbon store coupled to plant growth. Annu Rev Plant Biol, 71:217-245. [48]ThakurR, SoodA, NagarPK, et al., 2006. Regulation of growth of Lilium plantlets in liquid medium by application of paclobutrazol or ancymidol, for its amenability in a bioreactor system: growth parameters. Plant Cell Rep, 25(5):382-391. [49]ThompsonJD, GibsonTJ, PlewniakF, et al., 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res, 25(24):4876-4882. [50]van HarsselaarJK, LorenzJ, SenningM, et al., 2017. Genome-wide analysis of starch metabolism genes in potato (Solanum tuberosum L.). BMC Genomics, 18:37. [51]van TuylJM, ArensP, RamannaMS, et al., 2011. Lilium. In: Kole C (Ed.), Wild Crop Relatives: Genomic and Breeding Resources. Springer, Berlin Heidelberg, Germany, p.161-183. [52]van TuylJM, ArensP, ShahinA, et al., 2018. Lilium. In: van Huylenbroeck J (Ed.), Ornamental Crops. Springer, Cham, Switzerland, p.481-512. [53]WangDP, ZhangYB, ZhangZ, et al., 2010. KaKs_Calculator 2.0: a toolkit incorporating gamma-series methods and sliding window strategies. Genom Proteom Bioinf, 8(1):77-80. [54]WangX, FengB, XuZB, et al., 2014. Identification and characterization of granule bound starch synthase I (GBSSI) gene of tartary buckwheat (Fagopyrum tataricum Gaertn.). Gene, 534(2):229-235. [55]WuY, XiaYP, ZhangJP, et al., 2016. Low humic acids promote in vitro lily bulblet enlargement by enhancing roots growth and carbohydrate metabolism. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 17(11):892-904. [56]WuY, LiY, MaYD, et al., 2017. Hormone and antioxidant responses of Lilium Oriental hybrids ‘Sorbonne’ bulblets to humic acid treatments in vitro. J Hortic Sci Biotech, 92(2):155-167. [57]WuY, SunMY, ZhangJP, et al., 2019a. Differential effects of paclobutrazol on the bulblet growth of oriental lily cultured in vitro: growth behavior, carbohydrate metabolism, and antioxidant capacity. J Plant Growth Regul, 38(2):359-372. [58]WuY, MaYD, LiY, et al., 2019b. Plantlet regeneration from primary callus cultures of Lilium brownii F.E.Br. ex Miellez var. giganteum G. Y. Li & Z. H. Chen, a rare bulbous germplasm. In Vitro Cell Dev-Biol-Plant, 55(1):44-59. [59]XuH, YangPP, CaoYW, et al., 2020. Cloning and functional characterization of a flavonoid transport-related MATE gene in asiatic hybrid lilies (Lilium spp.). Genes, 11(4):418. [60]YangM, ZhuLP, PanC, et al., 2015. Transcriptomic analysis of the regulation of rhizome formation in temperate and tropical lotus (Nelumbo nucifera). Sci Rep, 5:13059. [61]YangPP, XuLF, XuH, et al., 2017. Histological and transcriptomic analysis during bulbil formation in Lilium lancifolium. Front Plant Sci, 8:1508. [62]ZhouYX, ChenYX, TaoX, et al., 2016. Isolation and characterization of cDNAs and genomic DNAs encoding ADP-glucose pyrophosphorylase large and small subunits from sweet potato. Mol Genet Genomics, 291(2):609-620. [63]ZhuFL, ChengN, SunH, et al., 2020. 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