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Somatic embryogenesis and plant regeneration in Piper aduncum L
In Vitro Cellular & Developmental Biology - Plant ( IF 2.2 ) Pub Date : 2020-08-28 , DOI: 10.1007/s11627-020-10110-y
Paulo Cesar Alves De Sousa , Stênio Steferson Silva E. Souza , Filipe Sathler Meira , Rennan De Oliveira Meira , Hugo Teixeira Gomes , Inaê Mariê De Araújo Silva-Cardoso , Jonny Everson Scherwinski-Pereira

An efficient protocol is reported for in vitro plant regeneration through somatic embryogenesis in Piper aduncum, a Brazilian Amazon species with high economic potential. The species is important due to a variety of components found in its essential oil, with emphasis on dillapiole. Leaf explants from five accessions identified for high oil yield and levels of dillapiole were evaluated for their embryogenic potential. To induce embryogenic calli, the explants were cultivated in MS medium supplemented with 5 mg L−1 of 1-naphthaleneacetic acid (NAA) and 2.5 mg L−1 of N6-benzylaminopurine (BAP) for 80 d. For somatic embryogenesis, the embryogenic calli were transferred to MS medium with 10 mg L−1 of NAA and 2.5 mg L1 of BAP and incubated for 45 d. The obtained somatic embryos were germinated in MS medium without regulators by 45 d and the obtained plantlets were subjected to acclimatization. Somatic embryos and calli from this process were subjected to anatomical and histochemical analyses. Biochemical analyses (total soluble sugars, starch, total amino acids, and proteins) were also performed to identify markers for embryogenic competence acquisition. In addition, the germination of somatic embryos was evaluated in a semi-solid and liquid system (R.I.T.A.® temporary immersion bioreactors). The obtained plants were evaluated for genetic fidelity using ISSR markers. The present study indicate that the accessions did not differ in embryogenic potential, with a mean percentage of calli with somatic embryos of 82.4%. Anatomical analyses confirmed the occurrence of the embryogenic route and the histochemical analyses identified starch grains in somatic embryos at different developmental stages. The biochemical analyses showed high total soluble sugars and total amino acids in embryogenic calli, marks of the acquisition of the embryogenic competence of P. aducum. The R.I.T.A.® temporary immersion bioreactors were highly efficient in the regeneration of somatic plants, with 100% germination. The plants regenerated in the semi-solid and liquid systems showed high genetic homogeneity. The survival rate of the acclimatized plants was 100%.



中文翻译:

东北胡椒的体细胞胚发生和植物再生

据报道,巴西亚马逊种具有很高的经济潜力的北美胡椒种中,通过体细胞胚发生进行体外植物再生的有效方案。由于其精油中发现了多种成分,因此该物种非常重要,其中以地那香酚为重点。评估了五个已鉴定出的高油产量和莳萝双酚水平的种的叶片外植体的胚发生潜力。为了诱导胚性愈伤组织,将外植体在补充有5 mg L -1的1-萘乙酸(NAA)和2.5 mg L -1的N6-苄基氨基嘌呤(BAP)的MS培养基中培养80 d。对于体细胞胚发生,将胚发生的愈伤组织转移至含10 mg L -1的MS培养基中NAA和2.5 mg L 1BAP并温育45天。将获得的体细胞胚在无调节剂的MS培养基中萌发45天,并使获得的小植株适应环境。对来自该过程的体细胞胚和愈伤组织进行了解剖和组织化学分析。还进行了生化分析(总可溶性糖,淀粉,总氨基酸和蛋白质)以鉴定用于胚胎发生能力获取的标志物。另外,在半固体和液体系统(RITA®临时浸入式生物反应器)中评估了体细胞胚的萌发。使用ISSR标记评估获得的植物的遗传保真度。本研究表明,这些种在胚发生潜能上没有差异,体细胞胚愈伤组织的平均百分比为82.4%。解剖学分析证实了胚发生途径的发生,并且组织化学分析确定了在不同发育阶段的体细胞胚中的淀粉粒。生化分析表明,胚性愈伤组织中可溶性总糖和氨基酸的含量高,这是获得胚性愈伤组织的标志。体育。RITA®临时浸入式生物反应器在体细胞再生中非常高效,发芽率为100%。在半固态和液态系统中再生的植物显示出很高的遗传同质性。适应植物的存活率为100%。

更新日期:2020-08-29
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