Laboratory of molecular biology of tomato, Bioengineering College, Chongqing University, Chongqing, 400044, People's Republic of China.
Sci Rep. 2017 Jul 19;7(1):5786. doi: 10.1038/s41598-017-04092-y.
The basic helix-loop-helix (bHLH) proteins are a large family of transcription factors that control various developmental processes in eukaryotes, but the biological roles of most bHLH proteins are not very clear, especially in tomato. In this study, a PRE-like atypical bHLH gene was isolated and designated as SlPRE2 in tomato. SlPRE2 was highly expressed in immature-green fruits, moderately in young leaves, flowers, and mature-green fruits. To further research the function of SlPRE2, a 35 S:PRE2 binary vector was constructed and transformed into wild type tomato. The transgenic plants showed increased leaf angle and stem internode length, rolling leaves with decreased chlorophyll content. The water loss rate of detached leaves was increased in young transgenic lines but depressed in mature leaves. Besides, overexpression of SlPRE2 promoted morphogenesis in seedling development, producing light-green unripening fruits and yellowing ripen fruits with reduced chlorophyll and carotenoid accumulation in pericarps, respectively. Quantitative RT-PCR analysis showed that expression of the chlorophyll related genes, such as GOLDEN 2-LIKE and RbcS, were decreased in unripening 35 S:PRE2 fruit, and carotenoid biosynthesis-related genes PHYTOENE SYNTHASE1A and ζ-CAROTENE DESATURASE in ripening fruit were also down-regulated. These results suggest that SlPRE2 affects plant morphology and is a negative regulator of fruit pigment accumulation.
碱性螺旋-环-螺旋(bHLH)蛋白是一类调控真核生物多种发育过程的转录因子家族,但大多数 bHLH 蛋白的生物学功能尚不清楚,尤其是在番茄中。本研究从番茄中分离并鉴定了一个类似 PRE 的非典型 bHLH 基因,命名为 SlPRE2。SlPRE2 在未成熟绿果中高表达,在幼叶、花和成熟绿果中中度表达。为进一步研究 SlPRE2 的功能,构建了 35S:PRE2 双元载体并转化野生型番茄。转基因植株表现出叶片角度增大、茎节间伸长、叶片卷曲、叶绿素含量降低。幼龄转基因植株的离体叶片失水率增加,而成熟叶片的失水率降低。此外,SlPRE2 的过表达促进了幼苗发育的形态发生,产生淡绿色的未成熟果实和黄色成熟果实,果皮中叶绿素和类胡萝卜素积累减少。定量 RT-PCR 分析表明,未成熟 35S:PRE2 果实中叶绿素相关基因如 GOLDEN 2-LIKE 和 RbcS 的表达降低,成熟果实中类胡萝卜素生物合成相关基因 PHYTOENE SYNTHASE1A 和 ζ-CAROTENE DESATURASE 的表达也下调。这些结果表明 SlPRE2 影响植物形态,是果实色素积累的负调控因子。