Cheng Jun, Shao Yun, Hu Xinyue, Gao Liying, Zheng Xianbo, Tan Bin, Ye Xia, Wang Wei, Zhang Haipeng, Wang Xiaobei, Lian Xiaodong, Li Zhiqian, Feng Jiancan, Zhang Langlang
Hortic Res. 2024 Jun 3;11(7):uhae155. doi: 10.1093/hr/uhae155. eCollection 2024 Jul.
Stable genetic transformation of peach [ (L.) Batsch] still faces many technical challenges, and existing transient expression methods are limited by tissue type or developmental stage, making it difficult to conduct functional analysis of genes regulating shoot growth. To overcome this dilemma, we developed a three-step method for efficient analysis of gene functions during peach seedling growth and development. This method resulted in transformation frequencies ranging from 48 to 87%, depending on the gene. From transformation of germinating seeds to phenotyping of young saplings took just 1.5 months and can be carried out any time of year. To test the applicability of this method, the function of three tree architecture-related genes, namely , , and , and two lateral root-related genes, and , were confirmed. Since functional redundancy can challenge gene functional analyses, tests were undertaken with the growth-repressor , which has three homologous genes, (), (), and (), in peach that are functionally redundant. Silencing using a triple-target vector (TRV2---) resulted in transgenic plants taller than those carrying just TRV2- or TRV2. Simultaneously silencing the three genes also attenuated the stature of two dwarf genotypes, 'FHSXT' and 'HSX', which normally accumulate DELLA proteins. Our study provides a method for the functional analysis of genes in peach and can be used for the study of root, stem, and leaf development. We believe this method can be replicated in other woody plants.
桃[ (L.) Batsch]的稳定遗传转化仍面临许多技术挑战，现有的瞬时表达方法受组织类型或发育阶段限制，难以对调控枝条生长的基因进行功能分析。为克服这一困境，我们开发了一种三步法，用于高效分析桃幼苗生长发育过程中的基因功能。该方法的转化频率在48%至87%之间，具体取决于基因。从种子萌发转化到幼树表型分析仅需1.5个月，且可在一年中的任何时间进行。为测试该方法的适用性，我们证实了三个与树形结构相关的基因，即 、 和 ，以及两个与侧根相关的基因 和 的功能。由于功能冗余会对基因功能分析造成挑战，我们对生长抑制因子 进行了测试，它在桃中有三个功能冗余的同源基因，即 ()、 ()和 ()。使用三靶点载体(TRV2---)进行沉默，得到的转基因植株比仅携带TRV2-或TRV2的植株更高。同时沉默这三个 基因也使两个通常积累DELLA蛋白的矮化基因型‘FHSXT’和‘HSX’的株高降低。我们的研究提供了一种对桃基因进行功能分析的方法，可用于研究根、茎和叶的发育。我们相信这种方法可以在其他木本植物中复制。
