Floral Transcriptome Sequencing for SSR Marker Development and Linkage Map Construction in the Tea Plant (Camellia sinensis)
Liqiang TanLiyuan WangKang WeiChengcai ZhangLiyun WuGui-Nian QiHao ChengQiang ZhangQingmei CuiJinbo Liang
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Abstract:
Despite the worldwide consumption and high economic importance of tea, the plant (Camellia sinensis) is not well studied in molecular biology. Under the few circumstances in which the plant is studied, C. sinensis flowers, which are important for reproduction and cross-breeding, receive less emphasis than investigation of its leaves or roots. Using high-throughput Illumina RNA sequencing, we analyzed a C. sinensis floral transcriptome, and 26.9 million clean reads were assembled into 75,531 unigenes averaging 402 bp. Among them, 50,792 (67.2%) unigenes were annotated with a BLAST search against the NCBI Non-Redundant (NR) database and 10,290 (16.67%) were detected that contained one or more simple sequence repeats (SSRs). From these SSR-containing sequences, 2,439 candidate SSR markers were developed and 720 were experimentally tested, validating 431 (59.9%) novel polymorphic SSR markers for C. sinensis. Then, a consensus SSR-based linkage map was constructed that covered 1,156.9 cM with 237 SSR markers distributed in 15 linkage groups. Both transcriptome information and the genetic map of C. sinensis presented here offer a valuable foundation for molecular biology investigations such as functional gene isolation, quantitative trait loci mapping, and marker-assisted selection breeding in this important species.Keywords:
Molecular breeding
Candidate gene
Illumina dye sequencing
Genetic linkage
茶樹の窒素要求性を解明する目的で,幼茶樹の窒素吸収能力の季節変化を調べた。気温の変化が露地に近い雨よけハウス内で,土耕栽培の鉢植え茶樹に毎月一定量の重窒素(15N)を与え,1ヶ月毎の15N吸収量を測定し,窒素吸収能力の季節変化を解析した。また,月別施用窒素の一番茶新芽窒素への寄与率を調査した。窒素吸収能力は春期から夏期にかけて高まり,8月に最高値に達した。その後徐々に低下し始め,厳冬期の2月に最低値を示した。2月の吸収能力は8月の約30%であった。3月以降,吸収能力は再び上昇を始めた。この窒素吸収能力の季節変動は気温変化と極めて類似していたことから,常緑樹である茶樹の養分吸収能力は,生育気温に大きく影響されるものと考えられた。一番茶新芽窒素に対する寄与率は,前年10月以降に施用された窒素が高く,特に当年春期3～4月の施用窒素が高かった。しかし,前年夏期以前に施用された窒素の寄与率も無視できない程高かった。
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The recent development of novel repeat-fruiting types of blackberry (Rubus L.) cultivars, combined with a long history of morphological marker-assisted selection for thornlessness by blackberry breeders, has given rise to increased interest in using molecular markers to facilitate blackberry breeding. Yet no genetic maps, molecular markers, or even sequences exist specifically for cultivated blackberry. The purpose of this study is to begin development of these tools by generating and annotating the first blackberry expressed sequence tag (EST) library, designing primers from the ESTs to amplify regions containing simple sequence repeats (SSR), and testing the usefulness of a subset of the EST-SSRs with two blackberry cultivars.A cDNA library of 18,432 clones was generated from expanding leaf tissue of the cultivar Merton Thornless, a progenitor of many thornless commercial cultivars. Among the most abundantly expressed of the 3,000 genes annotated were those involved with energy, cell structure, and defense. From individual sequences containing SSRs, 673 primer pairs were designed. Of a randomly chosen set of 33 primer pairs tested with two blackberry cultivars, 10 detected an average of 1.9 polymorphic PCR products.This rate predicts that this library may yield as many as 940 SSR primer pairs detecting 1,786 polymorphisms. This may be sufficient to generate a genetic map that can be used to associate molecular markers with phenotypic traits, making possible molecular marker-assisted breeding to compliment existing morphological marker-assisted breeding in blackberry.
Primer (cosmetics)
Rubus
Molecular breeding
Molecular marker
Marker-Assisted Selection
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Рассматриваются вопросы образования различных классов фенольных соединений в растениях чая (Camellia sinensis L.) и полученных из него клеточных культур. Сообщается об изменениях в их составе в условиях in vitro. Приводятся данные о регулирующем действии света на накопление фенольных соединений. The formation of various classes of phenolic compounds in tea plants (Camellia sinensis L.) and cell cultures derived from it has been reported. Provides information about the changes in their structure under in vitro. Data are presented on the regulatory effect of light on the accumulation of phenolic compounds.
Camellia
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Theaceae
Camellia
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Camellia
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Camellia
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Calmodulin-like (CML) proteins are a class of important Ca2+ sensors in plants, which play vital roles in regulating plant growth and development and response to abiotic stress. Tea plant (Camellia sinensis L.) is the most popular non-alcoholic economic beverage crop around the world. However, the potential functions of CMLs in either tea plants growth or in the response to environmental stresses are still unclear. In the present study, five CsCML genes (CsCML16, CsCML18-1, CsCML18-2, CsCML38, and CsCML42) were isolated from tea plant, and functionally characterized. The CsCML genes showed diverse expression patterns in leaves, roots, old stems, immature stems and flowers of tea plants. To investigate the expression changes of the genes under various abiotic stresses and ABA treatment, time-course experiments were also performed, the results indicated that the expression levels of CsCML16, 18-2 and 42 were significantly induced under low temperature and salt condition, while CsCML38 was induced distinctly under drought stress and ABA treatment. Overall, CsCML genes showed diverse function in tea plant under various stimuli. These results will increase our knowledge of the significance of CsCML genes in tea plant in response to abiotic stresses and hormone treatments.
Camellia
Plant Physiology
Theaceae
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