Wu Yongzhong, Fox Tim W, Trimnell Mary R, Wang Lijuan, Xu Rui-Ji, Cigan A Mark, Huffman Gary A, Garnaat Carl W, Hershey Howard, Albertsen Marc C
DuPont Pioneer, Johnston, IA, USA.
Plant Biotechnol J. 2016 Mar;14(3):1046-54. doi: 10.1111/pbi.12477. Epub 2015 Oct 6.
We have developed a novel hybridization platform that utilizes nuclear male sterility to produce hybrids in maize and other cross-pollinating crops. A key component of this platform is a process termed Seed Production Technology (SPT). This process incorporates a transgenic SPT maintainer line capable of propagating nontransgenic nuclear male-sterile lines for use as female parents in hybrid production. The maize SPT maintainer line is a homozygous recessive male sterile transformed with a SPT construct containing (i) a complementary wild-type male fertility gene to restore fertility, (ii) an α-amylase gene to disrupt pollination and (iii) a seed colour marker gene. The sporophytic wild-type allele complements the recessive mutation, enabling the development of pollen grains, all of which carry the recessive allele but with only half carrying the SPT transgenes. Pollen grains with the SPT transgenes exhibit starch depletion resulting from expression of α-amylase and are unable to germinate. Pollen grains that do not carry the SPT transgenes are nontransgenic and are able to fertilize homozygous mutant plants, resulting in nontransgenic male-sterile progeny for use as female parents. Because transgenic SPT maintainer seeds express a red fluorescent protein, they can be detected and efficiently separated from seeds that do not contain the SPT transgenes by mechanical colour sorting. The SPT process has the potential to replace current approaches to pollen control in commercial maize hybrid seed production. It also has important applications for other cross-pollinating crops where it can unlock the potential for greater hybrid productivity through expanding the parental germplasm pool.
我们开发了一种新型杂交平台,该平台利用核雄性不育在玉米和其他异花授粉作物中生产杂交种。该平台的一个关键组成部分是一个称为种子生产技术(SPT)的过程。这个过程包含一个转基因SPT保持系,它能够繁殖非转基因核雄性不育系,用作杂交种生产中的母本。玉米SPT保持系是一个纯合隐性雄性不育系,用一个包含(i)一个互补的野生型雄性育性基因以恢复育性,(ii)一个α-淀粉酶基因以破坏授粉,以及(iii)一个种子颜色标记基因的SPT构建体进行转化。孢子体野生型等位基因补充隐性突变,使花粉粒得以发育,所有花粉粒都携带隐性等位基因,但只有一半携带SPT转基因。携带SPT转基因的花粉粒由于α-淀粉酶的表达而出现淀粉耗尽,无法萌发。不携带SPT转基因的花粉粒是非转基因的,能够使纯合突变体植株受精,产生用作母本的非转基因雄性不育后代。由于转基因SPT保持系种子表达红色荧光蛋白,它们可以通过机械颜色分选被检测到并与不含SPT转基因的种子有效分离。SPT过程有可能取代商业玉米杂交种子生产中目前控制花粉的方法。它对其他异花授粉作物也有重要应用,通过扩大亲本种质库,它可以释放出提高杂交种生产力的潜力。