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An optimized procedure for plant recovery from somatic embryos significantly facilitates the genetic improvement of Vitis.

ABSTRACT: Plant regeneration from grapevine (Vitis spp.) via somatic embryogenesis typically is poor. Recovery of plants from Vitis rotundifolia Michx. (muscadine grape) is particularly problematic due to extremely low efficiency, including extended culture durations required for embryo-plant conversion. Poor plant recovery is an obstacle to the selection of improved genetically modified lines. Somatic embryos (SEs) of V. rotundifolia cultivar Delicious (Del-HS) and Vitis vinifera L cultivar Thompson Seedless (TS) were used to identify culture media and conditions that promoted embryo differentiation and plant conversion; this resulted in a two-step culture system. In comparative culture experiments, C2D medium containing 6% sucrose was the most effective, among four distinct formulae tested, for inducing precocious SE germination and cell differentiation. This medium, further supplemented with 4?µM 6-benzylaminopurine (C2D4B), was subsequently determined to enhance post-germinative growth of SE. MS medium supplemented with 0.5?µM 1-naphthaleneacetic acid (MSN) was then utilized to stimulate root and shoot growth of germinated SE. An average of 35% and 80% 'Del-HS' and 'TS' SE, respectively, developed into plants. All plants developed robust root and shoot systems and exhibited excellent survival following transfer to soil. Over 150 plants of 'Del-HS' were regenerated and established within 2.5?months, which is a dramatic reduction from the 6- to 12-month time period previously required. Similarly, 88 'TS' plant lines were obtained within the same time period. Subsequently, seven out of eight Vitis cultivars exhibited significantly increased plant conversion percentages, demonstrating broad application of the two-step culture system to produce the large numbers of independent plant lines needed for selection of desired traits.

SUBMITTER: Li ZT

PROVIDER: S-EPMC4596318 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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An optimized procedure for plant recovery from somatic embryos significantly facilitates the genetic improvement of Vitis.

Li Zhijian T ZT Kim Kyung-Hee KH Dhekney Sadanand A SA Jasinski Jonathan R JR Creech Matthew R MR Gray Dennis J DJ

Horticulture research 20140611

Plant regeneration from grapevine (Vitis spp.) via somatic embryogenesis typically is poor. Recovery of plants from Vitis rotundifolia Michx. (muscadine grape) is particularly problematic due to extremely low efficiency, including extended culture durations required for embryo-plant conversion. Poor plant recovery is an obstacle to the selection of improved genetically modified lines. Somatic embryos (SEs) of V. rotundifolia cultivar Delicious (Del-HS) and Vitis vinifera L cultivar Thompson Seed ...[more]

PMID: 26504540

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Project description:BackgroundSomatic embryogenesis in grapevines is a complex process that depends on many physiological and genetic factors. One of its main limitations is the process of precocious germination of the somatic embryos in differentiation medium. This process lowers plant conversion rates from the somatic embryos, and it is probably caused by a low endogenous abscisic acid (ABA) content.ResultsPrecocious germination of the somatic embryos was successfully avoided by culturing grapevine cv. Mencía embryogenic aggregates over a semipermeable membrane extended on top of the differentiation medium. The weekly analysis of the endogenous ABA and ABA-glucosyl ester (ABA-GE) contents in the aggregates showed their rapid accumulation. The expression profiles of 9-cis-epoxycarotenoid dioxygenase (VvNCED1), 8'-hydroxylase (VvHyd2), UDP-glucosyltransferase (VvUGT) and β-glucosidase (VvBG2) genes in grapevine revealed that the occurrence of a first accumulation peak of endogenous ABA in the second week of culture over the semipermeable membrane was mainly dependent on the expression of the VvNCED1 gene. A second increase in the endogenous ABA content was observed in the fourth week of culture. At this point in the culture, our results suggest that of those genes involved in ABA accumulation, one (VvNCED1) was repressed, while another (VvBG2) was activated. Similarly, of those genes related to a reduction in ABA levels, one (VvUGT) was repressed while another (VvHyd2) was activated. The relative expression level of the VvNCED1 gene in embryogenic aggregates cultured under the same conditions and treated with exogenous ABA revealed the significant downregulation of this gene.ConclusionsOur results demonstrated the involvement of ABA metabolism in the control of the maturation of grapevine somatic embryos cultured over a semipermeable membrane and two important control points for their endogenous ABA levels. Thus, subtle differences in the expression of the antagonistic genes that control ABA synthesis and degradation could be responsible for the final level of ABA during the maturation of grapevine somatic embryos in vitro. In addition, the treatment of somatic embryos with exogenous ABA suggested the feedback-based control of the expression of the VvNCED1 gene by ABA during the maturation of grapevine somatic embryos.

Project description:BackroundIn recent years, with the increasing demand for seedless grape varieties that have lower production costs, are disease resistant/tolerant and require less chemical pesticides, the embryo recovery technique has begun to be used more in table grape breeding studies. However, the desired high success rate has not yet been achieved in these studies. Although there are different reasons for this, especially the grape varieties selected for cross-breeding and the timing of transferring the embryos to medium are among the most important reasons. In this study, focusing on these two important factors, the embryos obtained from different hybridization combinations were transferred to agar medium at different weeks for 4 years and the most successful combination and time were determined. In addition, seedless and large berry grape varieties and some seeded varieties that are resistant/tolerant to fungal diseases were selected as parents because they can provide resistance to disease infections in vitro and thus increase the success rate.ResultsThe results obtained from the study showed that the selected variety and combination significantly affected the success rate in embryo rescue. Especially in combinations with the 'Yalova Seedless' variety as the female parent, more successful results were obtained compared to combinations of other varieties. When 'Yalova Seedless' variety was pollinated with pollen of 'Red Globe', 'Muscat Bailey A' and 'Exalta' varieties, more seedlings were obtained with the help of embryo rescue. The results obtained over four years showed that the best sampling time after pollination was the eighth week and then the seventh week.ConclusionsAccording to the results obtained, it has been shown that the selected varieties and the sampling time significantly affect the success rate in embryo rescue studies. Therefore, higher success rates can be achieved in comprehensive breeding studies in which they will be included as pollinators, especially in different seeded varieties that are resistant to diseases and have larger berry size.

Project description:BackgroundIt is of great significance to explore a path for expedited recovery from thoracic surgery for patients undergoing minimally invasive lobectomy to ensure their rapid and smooth recovery and to conserve medical resources.MethodsWe analyzed 629 cases from the Department of Thoracic Surgery, First Affiliated Hospital of Soochow University from January 2018 to January 2020. According to the length of postoperative stay (LOS) and perioperative management, the 629 patients were divided into group A [routine management group (RMG)], group B [rapidly recovery group (RRG), LOS >72 h], and group C (RRG, LOS ≤72 h). The t-test and chi-square test were used to compare the postoperative complications (PC), chest tube indwelling time (CTIT), LOS, postoperative opioid dosages (POD), and total costs (TC) of the 3 groups.ResultsCompared with the RMG, the LOS, PC, CTIT, POD, and TC of the RRG were statistically significantly ameliorated (P<0.05). When compared with group A, the PC (18.9% vs. 38.8%), LOS (2.74±0.80 vs. 5.70±1.10 d), CTIT (46.1±18.5 vs. 123.6±34.8 h), and TC (¥51,517±7,217 vs. ¥65,781±8,200) of group C were all decreased. Compared with group A, group B had more preoperative complications, less CTIT, but no significant difference in LOS, PC, and TC during hospital stay. Compared with group B, group C had less preoperative complications, and reduced LOS, CTIT, and TC. The POD in group A was significantly higher than that in groups B and C.ConclusionsUse of an enhanced recover after surgery (ERAS) protocol can reduce the postoperative complications and shorten the LOS. Some high-risk patients cannot be discharged within 72 h after surgery, but their incidence of grade II postoperative lung complications can be decreased and they can benefit from ERAS. Single intercostal nerve block, COX-2 selective inhibitor, and removal of the chest tube as early as possible can improve the ERAS.KeywordsEnhanced recover after surgery (ERAS); video-assisted thoracic surgery (VATS); minimally invasive lobectomy.

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An optimized procedure for plant recovery from somatic embryos significantly facilitates the genetic improvement of Vitis.

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An optimized procedure for plant recovery from somatic embryos significantly facilitates the genetic improvement of Vitis.

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