Plant Genetics Research Unit, USDA-Agricultural Research Service, Columbia, MO 65211, USA.
Plant Science Division, University of Missouri, Columbia, MO 65211, USA.
Int J Mol Sci. 2020 Jul 5;21(13):4775. doi: 10.3390/ijms21134775.
High growth temperatures negatively affect soybean ( (L.) Merr) yields and seed quality. Soybean plants, heat stressed during seed development, produce seed that exhibit wrinkling, discoloration, poor seed germination, and have an increased potential for incidence of pathogen infection and an overall decrease in economic value. Soybean breeders have identified a heat stress tolerant exotic landrace genotype, which has been used in traditional hybridization to generate experimental genotypes, with improved seed yield and heat tolerance. Here, we have investigated the seed protein composition and ultrastructure of cotyledonary parenchyma cells of soybean genotypes that are either susceptible or tolerant to high growth temperatures. Biochemical analyses of seed proteins isolated from heat-tolerant and heat-sensitive genotypes produced under 28/22 °C (control), 36/24 °C (moderate), and 42/26 °C (extreme) day/night temperatures revealed that the accumulation in soybean seeds of lipoxygenase, the β-subunit of β-conglycinin, sucrose binding protein and Bowman-Birk protease inhibitor were negatively impacted by extreme heat stress in both genotypes, but these effects were less pronounced in the heat-tolerant genotype. Western blot analysis showed elevated accumulation of heat shock proteins (HSP70 and HSP17.6) in both lines in response to elevated temperatures during seed fill. Transmission electron microscopy showed that heat stress caused dramatic structural changes in the storage parenchyma cells. Extreme heat stress disrupted the structure and the membrane integrity of protein storage vacuoles, organelles that accumulate seed storage proteins. The detachment of the plasma membrane from the cell wall (plasmolysis) was commonly observed in the cells of the sensitive line. In contrast, these structural changes were less pronounced in the tolerant genotype, even under extreme heat stress, cells, for the most part, retained their structural integrity. The results of our study demonstrate the contrasting effects of heat stress on the seed protein composition and ultrastructural alterations that contribute to the tolerant genotype's ability to tolerate high temperatures during seed development.
高温会对大豆( (L.) Merr)的产量和种子质量产生负面影响。在种子发育过程中受热胁迫的大豆植株会产生皱缩、变色、发芽不良的种子,并且更容易受到病原体感染的影响,经济价值也会降低。大豆育种家已经鉴定出一种耐热的外来地方品种基因型,该基因型已被用于传统杂交,以产生具有更高种子产量和耐热性的实验基因型。在这里,我们研究了对高温敏感或耐受的大豆基因型的子叶薄壁组织细胞的种子蛋白组成和超微结构。在 28/22°C(对照)、36/24°C(中度)和 42/26°C(极端)日/夜温度下,从耐热和热敏基因型中分离的种子蛋白进行生化分析表明,在两种基因型中,脂氧合酶、β-伴大豆球蛋白的β亚基、蔗糖结合蛋白和 Bowman-Birk 蛋白酶抑制剂在极端高温下的积累受到负面影响,但在耐热基因型中,这些影响不太明显。Western blot 分析表明,在种子灌浆过程中,两条线都因温度升高而积累了热休克蛋白(HSP70 和 HSP17.6)。透射电子显微镜显示,热应激导致贮藏薄壁细胞的结构发生剧烈变化。极端热应激破坏了蛋白储存液泡的结构和膜完整性,液泡是积累种子储存蛋白的细胞器。质膜与细胞壁分离(质壁分离)在敏感系细胞中很常见。相比之下,在耐受基因型中,这些结构变化不太明显,即使在极端高温下,细胞在很大程度上仍保持其结构完整性。我们的研究结果表明,热应激对种子蛋白组成和超微结构改变的影响截然不同,这有助于耐受基因型在种子发育过程中耐受高温的能力。