Rosa persica Michx ex Juss.的体细胞胚胎发生诱导

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-06-17 DOI:10.1007/s11240-024-02805-6
Shahrzad Vaziee, Mostafa K. Sarmast, Farshid Ghaderi-far, Changquang Wang
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引用次数: 0

摘要

目前的研究旨在评估各种外植体(未成熟的子叶胚、下胚轴和子叶)、液体和琼脂凝固的 MS 培养基以及 2,4-D 和 BA 对蔷薇体细胞胚的诱导和发育的影响。虽然下胚轴和子叶外植体能产生胼胝体和初级球状胚,但在半强度液体和琼脂凝固的 MS 培养基中,只有未成熟的子叶外植体能产生成熟的体细胞胚。只有在琼脂凝固的 MS 培养基中才能产生间接的成熟体细胞胚,而在液体 MS 培养基中产生的直接胚和间接胚不能进一步发育成成熟胚。胚胎性胼胝体的诱导主要发生在添加了 0.3 mg L- 1 2,4-D 的半强度 MS 培养基上,或在半强度 MS 液体培养基和琼脂固体培养基上分别发生在添加了 0.09 mg L- 1 BA 和 0.3 mg l- 1 2,4-D 的半强度 MS 培养基上。在半强度的 MS 琼脂固化培养基和液体培养基上,未成熟的合子胚形成胚生茧的成功率分别为 57.14% 和 46% 。只有从分离的未成熟子实体外植体上才能诱导间接体细胞胚胎发生,在琼脂固化培养基和液体培养基上的成功率分别为 25.11% 和 9.64%。然而,在液体培养基和琼脂固化培养基中,诱导最高体细胞胚胎的 PGR 浓度是不同的。在液体培养基中,将外植体放在添加了 1 mg L- 1 BA 和 0.9 mg L- 1 2,4-D 的半强度 MS 培养基上培养四周后,再放入添加了 1 mg L- 1 BA 和 0.3 mg L- 1 2,4-D 的半强度 MS 培养基中进行亚培养,可获得最高的间接体细胞胚(每个外植体可获得 24.5 个胚)。在相同的 MS 培养基中添加 1 mg L- 1 的 BA 和 0.09 mg L- 1 的 2,4-D,可获得直接体细胞胚(每个外植体 4.3 个胚)。但是,没有一个体细胞胚能在液体培养基中进一步发芽。在琼脂固体培养基中,在诱导阶段将外植体放在添加了 0.3 mg L- 1 2,4-D 的半强度 MS 培养基中培养,然后将胚性胼胝体放在添加了 1 mg L- 1 BA 和 0.3 mg L- 1 2,4-D 的半强度 MS 培养基中进行亚培养,可获得最多的体细胞胚(每个外植体有 26 个体细胞胚)。无 PGR 和琼脂固化 MS 培养基中的体细胞胚发芽率最高(46.15%),这些体细胞胚来自 16 周龄的胚性胼胝体培养,主要在添加 0.3 mg L- 1 2,4-D 的半强度 MS 培养基中培养四周,然后在添加 1 mg L- 1 BA 和 0.3 mg L- 1 2,4-D 的半强度 MS 培养基中进行亚培养。获得的小植株的适应效率为 25%,在温室中不能旺盛生长。该实验的结果有助于在育种计划中利用野生蔷薇物种作为重要的遗传资源,以扩大新蔷薇栽培品种的可获得基因库。
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Somatic embryogenesis induction in Rosa persica Michx ex Juss.

The current research aimed to evaluate the impact of various explants (immature zygote embryos, hypocotyls, and cotyledons), the effects of liquid and agar solidified MS media, as well as 2,4-D and BA on the induction and development of somatic embryos from Rosa persica. Although hypocotyls and cotyledon explants generated callus and primary globular embryos, only immature zygotic explants were able to produce mature somatic embryos in a half-strength liquid and agar- solidified MS medium. Indirect mature somatic embryos were only produced in agar-solidified MS media while liquid MS media-derived direct and indirect embryos could not further develop into mature embryo. Embryogenic callus induction mostly occcurred in half-strength MS media supplemented with 0.3 mg L− 1 2,4-D or 0.09 mg L− 1 BA and 0.3 mg l− 1 2,4-D on half-strength MS liquid and agar-solidified media, respectively. Immature zygote embryos formed embryogenic calli at a success rate of 57.14% and 46% on half-strength MS agar-solidified and liquid media, respectively. Indirect Somatic embryogenesis was induced only from detached immature zygote explants, achieving a frequency of 25.11% and 9.64% in agar-solidified and liquid media, respectively. However, the concentration of PGR to induce the highest somatic embryo was different in liquid and agar-solidefied media. In liquid media, the incubation of explant for four weeks on half-strength MS media supplemented with 1 mg L− 1 of BA and 0.9 mg L− 1 2,4-D and then subculturing them in half-strength MS media supplemented with 1 mg L− 1 of BA and 0.3 mg L− 1 2,4-D resulted in the highest indirect somatic embryo (24.5 embryos per explant). The same MS media supplemented with 1 mg L− 1 of BA and 0.09 mg L− 1 2,4-D resulted in direct somatic embryo (4.3 embryos per explant). However, non of the somatic embryos could further germinate in liquid media. In agar-solidified media, the incubation of explants in half-strength MS media supplemented with 0.3 mg L− 1 2,4-D during the induction phase and subsequent subculture of embryogenic callus on half-strength MS media supplemented with 1 mg L− 1 of BA and 0.3 mg L− 1 2,4-D resulted in the highest somatic embryo (26 somatic embryos per explant). The highest germination rate (46.15%) of somatic embryos occurred in PGR-free and agar-solidified MS media from 16-weeks-old embryogenic callus culture, which was primarily incubated in half-strength MS media supplemented with 0.3 mg L− 1 2,4-D for four weeks and then subculturing them in half-strength MS media supplemented with 1 mg L− 1 of BA and 0.3 mg L− 1 2,4-D. The acclimatization efficiency of the acquired plantlets was 25%, and they could not grow vigorously in the greenhouse. The results of this experiment can help utilize wild relative Rosa species in breeding programs as a remarkable genetic resource to expand the accessible gene pool of new Rosa cultivars.

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ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
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2.10%
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464
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