Response of the transcription factor BABY BOOM of Arabidopsis thaliana L. in the formation of embryogenic calluses of cocoa leaves (Theobroma cacao L.)

IF 1 Q3 AGRICULTURE, DAIRY & ANIMAL SCIENCE
Nikkolas Javier Alburqueque-Vasquez, Angel David Hernández-Amasifuen, Alexandra Jherina Pineda-Lázaro, Jaime N. Fernández Ponce, Juan Carlos Guerrero-Abad, Ligia García, Mike Anderson Corazon-Guivin
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Abstract

Cocoa (Theobroma cacao L.) is one of the most important economic crops worldwide. The propagation of elite varieties of cocoa has been achieved through somatic embryogenesis, but still one of the main limitations is the low rates of embryo formation, which is a genotype-dependent trait. Manipulation of transcription factors (TFs) such as BABY BOOM (BBM) promotes the transition of cocoa somatic cells from the vegetative to the embryonic state. This work validated the use of clonal cocoa leaves cv. IMC-67 to induce somatic embryogenesis, overcoming their recalcitrant limitation with the help of the introduction of TF-BBM from Arabidopsis thaliana (AtBBM). The vectors were constructed by the Gateway system using the donor vector pENTR/D-TOPO and the expression vector pk7WG2. The overexpression vector pk7WG2:AtBBM was obtained, allowing successful transformation into Agrobacterium tumefaciens GV3101. The AtBBM gene was characterized (1755 base pairs), and its expression was observed in the formation of embryogenic calluses in cocoa leaves. Overexpression of AtBBM allowed the obtainment of a 92% response in the formation of embryogenic callus in cocoa leaves with Agrobacterium-mediated vacuum infiltration and overexpression of the pk7WG2:AtBBM vector. This high transformation efficiency reached with the insertion of the overexpression vector provides validation of transient response of the TF AtBBM in the formation of embryogenic calluses in cocoa leaves of the IMC-67 clone. Through this methodology, it is possible to continue with studies of gene overexpression, insertion, silencing, and gene editing in Peruvian cocoa.
拟南芥转录因子BABY BOOM对可可胚性愈伤组织形成的响应
可可(Theobroma Cocoa L.)是世界上最重要的经济作物之一。可可优良品种的繁殖是通过体细胞胚发生实现的,但主要的限制之一是胚胎形成率低,这是一个基因型依赖性性状。转录因子(tf)如BABY BOOM (BBM)的调控促进可可体细胞从营养状态向胚胎状态的转变。本工作验证了克隆可可叶cv的使用。通过引入拟南芥(Arabidopsis thaliana, AtBBM)的TF-BBM, IMC-67诱导体细胞胚胎发生,克服了其顽固性的限制。载体由Gateway系统构建,供体载体为pENTR/D-TOPO,表达载体为pk7WG2。获得过表达载体pk7WG2:AtBBM,成功转化为农杆菌GV3101。鉴定了AtBBM基因(1755个碱基对),并在可可胚性愈伤组织形成过程中观察到其表达。通过农杆菌介导的真空浸润和pk7WG2:AtBBM载体的过表达,AtBBM在可可叶片胚性愈伤组织形成中的应答率达到92%。通过插入过表达载体达到的高转化效率验证了TF AtBBM在IMC-67克隆可可叶片胚性愈伤组织形成中的瞬态响应。通过这种方法,可以继续研究秘鲁可可中的基因过表达、插入、沉默和基因编辑。
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来源期刊
Scientia Agropecuaria
Scientia Agropecuaria AGRICULTURE, DAIRY & ANIMAL SCIENCE-
CiteScore
3.50
自引率
0.00%
发文量
27
审稿时长
12 weeks
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