Department of Plant and Environmental Sciences, School of Health Research, Clemson University Pee Dee Research and Education Centre, Florence, SC, USA.
Department of Crop and Soil Sciences, Washington State University, Pullman, WA, USA.
Methods Mol Biol. 2020;2124:263-279. doi: 10.1007/978-1-0716-0356-7_14.
There are specific advantages of using microspores as explants: (1) A small number of explant donors are required to obtain the desired number of pollen embryoids for genetic transformation and (2) microspores constitute a synchronous mass of haploid cells, which are transformable by various means and convertible to doubled haploids therefore allow production of homozygous genotypes in a single generation. Additionally, it has been demonstrated in wheat and other crops that microspores can be easily induced to produce embryoids and biolistic approach to produce a large number of transformants. In view of these listed advantages, we optimized the use of microspore-derived calli for biolistic transformation of wheat. The procedure takes about 6 months to obtain the viable transformants in the spring wheat background. In the present communication, we demonstrated the use of this method to produce the reduced immunogenicity wheat genotypes.
（1）只需少量的外植体供体就能获得所需数量的花粉胚状体用于遗传转化，（2）小孢子是由大量同步的单倍体细胞组成，这些细胞可以通过各种手段转化，并可转化为加倍的单倍体，因此可以在一代中产生纯合基因型。此外，在小麦和其他作物中已经证明，小孢子很容易被诱导产生胚状体，而且通过弹道方法可以产生大量的转化体。鉴于这些列出的优势，我们优化了利用小孢子衍生的愈伤组织进行小麦的弹道转化。在春季小麦背景下，该过程大约需要 6 个月才能获得有活力的转化体。在本研究中，我们展示了这种方法在生产低免疫原性小麦基因型中的应用。
