High frequency adventitious shoot regeneration from hypocotyl-derived callus of Glyptostrobus pensilis, a critically endangered plant
High frequency adventitious shoot regeneration from hypocotyl-derived callus of Glyptostrobus pensilis, a critically endangered plant
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DOI：
10.1007/s11240-022-02396-0
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发表时间：
2022-10
期刊：
影响因子：
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通讯作者：
Lu Yuan;Siyu Ma;Kenneth K. M. Liu;Ting Wang;D. Xiao;A. Zhang;Bin Liu;Ligen Xu;Riquan Chen;Liping Chen
中科院分区：
文献类型：
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作者：
Lu Yuan;Siyu Ma;Kenneth K. M. Liu;Ting Wang;D. Xiao;A. Zhang;Bin Liu;Ligen Xu;Riquan Chen;Liping Chen
水松是世界上极度濒危的植物物种，在自然条件下发芽率低，再生能力差。本研究通过序列相关扩增多态性分析表明，采用种子培养进行有性繁殖可以提高该植物的遗传多样性。对不同发育阶段的种子进行了发芽试验。在10月份未成熟期取样的种子在离体条件下萌发率最高（19.2%），在半强度DCR基本培养基上培养时产生幼苗。随后，切下幼苗的下胚轴并在愈伤组织和芽诱导培养基（CSIM）上孵育，所述愈伤组织和芽诱导培养基（CSIM）包括补充有2 mg L-16-苄基腺嘌呤（6-BA）和0.5mg L-1α-萘乙酸（NAA）的Murashige和Skoog（MS）基础培养基。然后将下胚轴来源的愈伤组织用作芽再生的起始材料。平均而言，2.4不定芽可以从一个下胚轴来源的愈伤组织再生。此外，当在CSIM上继代时，从附着于下胚轴来源的部分愈伤组织的再生不定芽中获得了15个以上的不定芽。生根驯化60天后，再将其移栽到保护区，形成绿巨人林。从而建立了一个循环的、高效率的、具有良好再生能力的绿绒藜不定芽再生体系。以下胚轴愈伤组织为起始材料。
Glyptostrobus pensilis, a critically endangered plant species worldwide, exhibits low germination rate and consequently poor regeneration ability under natural conditions. In this study, sequence-related amplified polymorphism analysis shows that sexual propagation of this plant species using seeds culture could improve its genetic diversity. Germination trials of seeds at different developmental stages were performed. Seeds sampled at the immature stage in October showed the highest germination rate (19.2%) in vitro, and produced seedlings when cultured on half-strength DCR basal medium. Subsequently, the hypocotyls of seedlings were excised and incubated on callus and shoot induction medium (CSIM), comprising Murashige and Skoog (MS) basal medium supplemented with 2 mg L−16-benzyladenine (6-BA) and 0.5 mg L−1α-naphthaleneacetic acid (NAA). The hypocotyl-derived callus was then used as starting material for shoot regeneration. On average, 2.4 adventitious shoots could be regenerated from a single hypocotyl-derived callus. Furthermore, more than 15 adventitious shoots were obtained from a regenerated adventitious shoot attached to a part of hypocotyl-derived callus, when sub-cultured on CSIM. After rooting and acclimation for 60 days, the plantlets were reintroduced into the conservation area, forming forests ofG. pensilis.Thus, we established a circular and highly efficient adventitious shoot regeneration system ofG. pensilisusing hypocotyl-derived callus as the starting material.