RESUMEN
Carbohydrates are important components in sweetpotatoes in terms of both their industrial use and eating quality. Although there has been a narrow range of diversity in the properties of sweetpotato starch, unique varieties and experimental lines with different starch traits have been produced recently both by conventional breeding and genetic engineering. The diversity in maltose content, free sugar composition and textural properties in sweetpotato cultivars is also important for their eating quality and processing of storage roots. In this review, we summarize the current status of research on and breeding for these important traits and discuss the future prospects for research in this area.
RESUMEN
Sweetpotato (Ipomoea batatas) is an autohexaploid species with 90 chromosomes (2n = 6x = 90) and a basic chromosome number of 15, and is therefore regarded as one of the most challenging species for high-density genetic map construction. Here, we used single nucleotide polymorphisms (SNPs) identified by double-digest restriction site-associated DNA sequencing based on next-generation sequencing technology to construct a map for sweetpotato. We then aligned the sequence reads onto the reference genome sequence of I. trifida, a likely diploid ancestor of sweetpotato, to detect SNPs. In addition, to simplify analysis of the complex genetic mode of autohexaploidy, we used an S1 mapping population derived from self-pollination of a single parent. As a result, 28,087 double-simplex SNPs showing a Mendelian segregation ratio in the S1 progeny could be mapped onto 96 linkage groups (LGs), covering a total distance of 33,020.4 cM. Based on the positions of the SNPs on the I. trifida genome, the LGs were classified into 15 groups, each with roughly six LGs and six small extra groups. The molecular genetic techniques used in this study are applicable to high-density mapping of other polyploid plant species, including important crops.
Asunto(s)
Genoma de Planta , Ipomoea batatas/genética , Polimorfismo de Nucleótido Simple , PoliploidíaRESUMEN
Ipomoea trifida (H. B. K.) G. Don. is the most likely diploid ancestor of the hexaploid sweet potato, I. batatas (L.) Lam. To assist in analysis of the sweet potato genome, de novo whole-genome sequencing was performed with two lines of I. trifida, namely the selfed line Mx23Hm and the highly heterozygous line 0431-1, using the Illumina HiSeq platform. We classified the sequences thus obtained as either 'core candidates' (common to the two lines) or 'line specific'. The total lengths of the assembled sequences of Mx23Hm (ITR_r1.0) was 513 Mb, while that of 0431-1 (ITRk_r1.0) was 712 Mb. Of the assembled sequences, 240 Mb (Mx23Hm) and 353 Mb (0431-1) were classified into core candidate sequences. A total of 62,407 (62.4 Mb) and 109,449 (87.2 Mb) putative genes were identified, respectively, in the genomes of Mx23Hm and 0431-1, of which 11,823 were derived from core sequences of Mx23Hm, while 28,831 were from the core candidate sequence of 0431-1. There were a total of 1,464,173 single-nucleotide polymorphisms and 16,682 copy number variations (CNVs) in the two assembled genomic sequences (under the condition of log2 ratio of >1 and CNV size >1,000 bases). The results presented here are expected to contribute to the progress of genomic and genetic studies of I. trifida, as well as studies of the sweet potato and the genus Ipomoea in general.
Asunto(s)
Variaciones en el Número de Copia de ADN , Genes de Plantas , Genoma de Planta , Ipomoea/genética , Secuencia de Bases , Genómica , Datos de Secuencia Molecular , Polimorfismo de Nucleótido Simple , Análisis de Secuencia de ADNRESUMEN
The sweetpotato cultivar Quick Sweet (QS) with a lower pasting temperature of starch is a unique breeding material, but the biochemical background of this property has been unknown. To assess the physiological impact of the reduced isoform II activity of starch synthase (SSII) on the starch properties in sweetpotato storage root, transgenic sweetpotato plants with reduced expressions of the SSII gene were generated and evaluated. All of the starches from transgenic plants showed lower pasting temperatures and breakdown measured by a Rapid Visco Analyzer. The pasting temperatures in transgenic plants were approximately 10-15 degrees C lower than in wild-type plants. Distribution of the amylopectin chain length of the transgenic lines showed marked differences compared to that in wild-type plants: more chains with degree of polymerization (DP) 6-11 and fewer chains with DP 13-25. The starch granules from the storage root of transgenic plants showed cracking on the hilum, while those from wild-type plants appeared to be typical sweetpotato starch. In accordance with these observations, the expression of SSII in the storage roots of the sweetpotato cultivar with low pasting temperature starch (QS) was notably lower than in cultivars with normal starch. Moreover, nucleotide sequence analysis suggested that most of the SSII transcripts in the cultivar with low pasting temperature starch were inactive alleles. These results clearly indicate that the activity of SSII in sweetpotato storage roots, like those in other plants, affects the pasting properties of starch through alteration of the amylopectin structure.
Asunto(s)
Amilopectina/química , Ipomoea batatas/genética , Proteínas de Plantas/metabolismo , Tubérculos de la Planta/enzimología , Almidón Sintasa/metabolismo , Temperatura , ADN Complementario/genética , Regulación de la Expresión Génica de las Plantas , Ipomoea batatas/enzimología , Isoenzimas/genética , Isoenzimas/metabolismo , Filogenia , Proteínas de Plantas/genética , Tubérculos de la Planta/genética , Plantas Modificadas Genéticamente/enzimología , Plantas Modificadas Genéticamente/genética , ARN de Planta/genética , Análisis de Secuencia de ADN , Almidón Sintasa/genéticaRESUMEN
In order to characterize the functions of the sweetpotato SRF1 gene, which encodes a Dof zinc finger transcriptional factor preferentially expressed in the storage roots, we isolated its full length cDNA and produced transgenic sweetpotato plants with altered SRF1 expression levels. The isolated cDNA of SRF1 encoded a polypeptide of 497 amino acids and was closely related to the cyclic Dof factors of Arabidopsis and the ascorbate oxidase binding protein of pumpkin. SRF1 was most highly expressed in storage roots, although some expression was also observed in other vegetative tissue. Transgenic plants overexpressing SRF1 showed significantly higher storage root dry matter content compared to the original cultivar Kokei No. 14 or control transgenic plants. In these plants, the starch content per fresh weight of the storage roots was also higher than that of the wild-type plants, while the glucose and fructose content drastically decreased. Among the enzymes involved in the sugar metabolism, soluble acid invertase showed a decreased activity in the transgenic plants. Gene expression analysis showed that the expression of Ibbetafruct2, which encodes an isoform of vacuolar invertase, was suppressed in the transgenic plants overexpressing the SRF1 gene. These data suggest that SRF1 modulates the carbohydrate metabolism in the storage roots through negative regulation of a vacuolar invertase gene.
Asunto(s)
Metabolismo de los Hidratos de Carbono/genética , Genes de Plantas , Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Raíces de Plantas/metabolismo , Factores de Transcripción/genética , Dedos de Zinc/genética , Secuencia de Aminoácidos , Biomasa , Regulación de la Expresión Génica de las Plantas , Ipomoea batatas/enzimología , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raíces de Plantas/anatomía & histología , Raíces de Plantas/genética , Plantas Modificadas Genéticamente , Solubilidad , Almidón/metabolismo , Factores de Transcripción/química , Factores de Transcripción/metabolismoRESUMEN
As a first step in clarifying the involvement of class I knotted1-like homeobox (KNOXI) genes in the storage root development of sweetpotato (Ipomoea batatas), we isolated three KNOXI genes, named Ibkn1, Ibkn2 and Ibkn3, expressed in the storage roots. Phylogenetic analysis showed that Ibkn1 was homologous to the SHOOT MERISTEMLESS (STM) gene of Arabidopsis, while Ibkn2 and Ibkn3 were homologous to the BREVIPEDICELLUS (BP) gene. Of these, expression of Ibkn1 and Ibkn2 were upregulated in developing and mature storage roots compared with fibrous roots. Ibkn1 and Ibkn2 showed different expression patterns in the storage roots. Ibkn1 was preferentially expressed at the proximal end and around the primary vascular cambium, while Ibkn2 expression was highest in the thickest part and lower in both the proximal and distal ends. In contrast to Ibkn1 and Ibkn2, expression of Ibkn3 in roots was not consistent among sweetpotato cultivars. The distribution of endogenous trans-zeatin riboside (t-ZR) in sweetpotato roots showed a similarity to the expression pattern of KNOXI genes, supporting the idea that KNOXI genes control cytokinin levels in the storage roots. The physiological functions of these KNOXI genes in storage root development are discussed.
Asunto(s)
Proteínas de Homeodominio/metabolismo , Ipomoea batatas/metabolismo , Proteínas de Plantas/metabolismo , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Citocininas/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Homeodominio/química , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/fisiología , Ipomoea batatas/genética , Ipomoea batatas/crecimiento & desarrollo , Isopenteniladenosina/análogos & derivados , Isopenteniladenosina/metabolismo , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/fisiología , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Alineación de SecuenciaRESUMEN
Granule-bound starch synthase I (GBSSI) is one of the key enzymes catalyzing the formation of amylose, a linear alpha(1,4)D-glucan polymer, from ADP-glucose. Amylose-free transgenic sweet potato plants were produced by inhibiting sweet potato GBSSI gene expression through RNA interference. The gene construct consisting of an inverted repeat of the first exon separated by intron 1 of GBSSI driven by the CaMV 35S promoter was integrated into the sweet potato genome by Agrobacterium tumefaciens-mediated transformation. In over 70% of the regenerated transgenic plants, the expression of GBSSI was inactivated giving rise to storage roots containing amylopectin but not amylose. Electrophoresis analysis failed to detect the GBSSI protein, suggesting that gene silencing of the GBSSI gene had occurred. These results clearly demonstrate that amylose synthesis is completely inhibited in storage roots of sweet potato plants by the constitutive production of the double-stranded RNA of GBSSI fragments. We conclude that RNA interference is an effective method for inhibiting gene expression in the starch metabolic pathway.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Ipomoea batatas/enzimología , Ipomoea batatas/genética , Interferencia de ARN , Almidón Sintasa/genética , Amilosa/análisis , Amilosa/metabolismo , Ipomoea batatas/metabolismo , Tubérculos de la Planta/genética , Tubérculos de la Planta/metabolismo , Plantas Modificadas Genéticamente , Almidón Sintasa/metabolismo , Transformación GenéticaRESUMEN
To identify the genes involved in storage root formation of sweet potato (Ipomoea batatas), we performed a simplified differential display analysis on adventitious roots at different developmental stages of the storage root. The expression patterns were confirmed by semiquantitative RT-PCR analyses. As a result, 10 genes were identified as being developmentally regulated and were named SRF1-SRF10. The expression of SRF1, SRF2, SRF3, SRF5, SRF6, SRF7, and SRF9 increased during storage root formation, whereas the expression of SRF4, SRF8, and SRF10 decreased. For further characterization, a full-length cDNA of SRF6 was isolated from the cDNA library of the storage root. SRF6 encoded a receptor-like kinase (RLK), which was structurally similar to the leucine-rich repeat (LRR) II RLK family of Arabidopsis thaliana. RNA gel blot analysis showed that the mRNA of SRF6 was most abundantly expressed in the storage roots, although a certain amount of expression was also observed in other vegetative organs. Tissue print mRNA blot analysis of the storage root showed that the mRNA of SRF6 was localized around the primary cambium and meristems in the xylem, which consist of actively dividing cells and cause the thickening of the storage root.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica/fisiología , Regulación de la Expresión Génica de las Plantas/fisiología , Ipomoea batatas/genética , Raíces de Plantas/metabolismo , Secuencia de Aminoácidos , Ipomoea batatas/crecimiento & desarrollo , Ipomoea batatas/metabolismo , Datos de Secuencia Molecular , Filogenia , Raíces de Plantas/crecimiento & desarrolloRESUMEN
A genomic fragment containing the dihydroflavonol 4-reductase B (DFR-B) gene was cloned from the sweet potato (Ipomoea batatas) and its nucleotide sequence was analyzed. The exons and flanking regions were highly homologous to those of previously reported DFR-B genes of the Japanese morning glory, whereas the introns and the intergenic region were less conserved. In addition to the sequences of three miniature inverted-repeat transposable elements (MITEs) and one direct repeat previously reported in the DFR-B gene of Japanese morning glory, two mobile element-like sequences were newly identified in the sweet potato DFR-B gene. At least four allelic sequences were found to exist by amplification of the DFR-B gene from various sweet potato cultivars. One of these allelic sequences had a 2-kb deletion in the intergenic region and was observed in the cultivars with high anthocyanin content in their storage roots.
Asunto(s)
Oxidorreductasas de Alcohol/genética , Ipomoea batatas/genética , Oxidorreductasas de Alcohol/metabolismo , Secuencia de Bases , Clonación Molecular , Elementos Transponibles de ADN , Dosificación de Gen , Ipomoea batatas/enzimología , Ipomoea batatas/metabolismo , Datos de Secuencia Molecular , Filogenia , Polimorfismo Genético , Análisis de Secuencia de ADNRESUMEN
To change the hexose-to-sucrose ratio within phloem cells, yeast-derived cytosolic invertase was expressed in transgenic potato (Solanum tuberosum cv. Desirée) plants under control of the rolC promoter. Vascular tissue specific expression of the transgene was verified by histochemical detection of invertase activity in tuber cross-sections. Vegetative growth and tuber yield of transgenic plants was unaltered as compared to wild-type plants. However, the sprout growth of stored tubers was much delayed, indicating impaired phloem-transport of sucrose towards the developing bud. Biochemical analysis of growing tubers revealed that, in contrast to sucrose levels, which rapidly declined in growing invertase-expressing tubers, hexose and starch levels remained unchanged as compared to wild-type controls. During storage, sucrose and starch content declined in wild-type tubers, whereas glucose and fructose levels remained unchanged. A similar response was found in transgenic tubers with the exception that starch degradation was accelerated and fructose levels increased slightly. Furthermore, changes in carbohydrate metabolism were accompanied by an elevated level of phosphorylated intermediates, and a stimulated rate of respiration. Considering that sucrose breakdown was restricted to phloem cells it is concluded that, in response to phloem-associated sucrose depletion or hexose elevation, starch degradation and respiration is triggered in parenchyma cells. To study further whether elevated hexose and/or hexose-phosphates or decreased sucrose levels are responsible for the metabolic changes observed, sucrose content was decreased by tuber-specific expression of a bacterial sucrose isomerase. Sucrose isomerase catalyses the reversible conversion of sucrose into palatinose, which is not further metabolizable by plant cells. Tubers harvested from these plants were found to accumulate high levels of palatinose at the expense of sucrose. In addition, starch content decreased slightly, while hexose levels remained unaltered, compared with the wild-type controls. Similar to low sucrose-containing invertase tubers, respiration and starch breakdown were found to be accelerated during storage in palatinose-accumulating potato tubers. In contrast to invertase transgenics, however, no accumulation of phosphorylated intermediates was observed. Therefore, it is concluded that sucrose depletion rather than increased hexose metabolism triggers reserve mobilization and respiration in stored potato tubers.