College of Bioresource Sciences, Nihon University, Kameino, Fujisawa, 252-0880, Japan.
Graduate School of Bioresource Sciences, Nihon University, Kameino, Fujisawa, 252-0880, Japan.
Physiol Plant. 2019 Oct;167(2):264-275. doi: 10.1111/ppl.12886. Epub 2019 Jan 9.
We investigated changes in starch content and starch metabolic enzyme activities in developing and postharvest squash of distinct species, Cucurbita maxima and Cucurbita moschata, which accumulate high and low levels of starch, respectively. The total activity of starch phosphorylase in developing fruits significantly correlated (r = 0.99) to the amount of starch among Cucurbita species (C. maxima, C. moschata and C. pepo). Separable activity of a plastidial L-form phosphorylase in C. maxima fruit markedly increased corresponding with starch accumulation. We isolated two genes (CmPhoL1 and CmPhoH1) encoding an L-form and a cytosolic H-form phosphorylase from C. maxima fruit. The expression of CmPhoL1 in the fruit dramatically increased at the beginning of starch accumulation. Recombinant CmPhoL1 enzyme showed similar kinetic parameters in both glucan synthesis and phosphorolysis: this enzyme can catalyze the invertible reaction in vitro depending on the concentration of substrates. These results suggest that CmPhoL1 plays a role in the starch accumulation process during squash development, but the aid of other starch synthetic enzymes may be required for in vivo glucan synthesis reaction by CmPhoL1. An importance of plastidial starch phosphorylase in the starch accumulation in the fruit organ was indicated.
我们研究了不同种南瓜(分别是高淀粉含量的 Cucurbita maxima 和低淀粉含量的 Cucurbita moschata)在发育中和采后淀粉含量和淀粉代谢酶活性的变化。发育中的果实中淀粉磷酸化酶的总活性与不同南瓜属物种(C. maxima、C. moschata 和 C. pepo)之间的淀粉含量呈显著相关(r = 0.99)。C. maxima 果实中质体 L 型磷酸化酶的可分离活性随着淀粉的积累而显著增加。我们从 C. maxima 果实中分离到两个编码 L 型和胞质 H 型磷酸化酶的基因(CmPhoL1 和 CmPhoH1)。在淀粉积累初期,CmPhoL1 在果实中的表达显著增加。重组 CmPhoL1 酶在葡聚糖合成和磷酸解中表现出相似的动力学参数:该酶可以根据底物浓度在体外催化可逆反应。这些结果表明,CmPhoL1 在南瓜发育过程中的淀粉积累过程中起作用,但 CmPhoL1 可能需要其他淀粉合成酶的辅助才能进行体内葡聚糖合成反应。这表明质体淀粉磷酸化酶在果实器官的淀粉积累中具有重要作用。