Deciphering transcriptional programming during lupin (Lupinus angustifolius) seed development using RNA-seq

VIRGINIA WAINAINA, Tina Rathjen, Trijntje Hughes, Annelie Marquardt, Natalie Fletcher, Hayley Casarotto, Meredith McNeil, Kerensa McElroy, Ling-Ling Gao
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Abstract

Lupin (Lupinus spp.) seeds are valued for their high protein content (35-40%) for both human and animal consumption. Seed development in crop plants is a critical factor influencing both seed fate and yield, hence, understanding the molecular mechanisms of seed development is essential. This study conducted a transcriptome analysis of Narrow Leaf Lupin (NLL) during seed development stages (3, 6, 9, 12, 15, 18, and 21 days after flowering) to investigate transcriptional dynamics and identify key candidate genes that control seed development. Approximately 357 million sequencing reads were generated from nine samples from leave, flower and seed tissues, enabling the identification of 34,769 expressed genes. The analysis revealed dynamic gene expression, with early stages marked by high metabolic activity and later stages focusing on storage protein synthesis and nutrient reservoir formation. The differential expression patterns of seed storage protein genes, including cupin groups (α, β, γ, and δ conglutins), were notable. The expression of α and β conglutins increased at later stages (15-21 days after flowering), supporting their role in grain filling and nutrient storage. Genes related to quinolizidine alkaloid biosynthesis, such as lysine/ornithine decarboxylase and purine permease transporter 1, showed late expression patterns suggesting alkaloid synthesis and transport during later stages. Many of the well-established transcription factors (TFs) known for their roles in seed development (bHLH, AP2, MYB, ERF, C2H2, NAC, WRKY, and C3H zinc finger families) showed differential expression, thus reinforcing the validity of our findings. These findings lay the groundwork for understanding the genetic and molecular mechanisms of seed development in lupin, contributing to enhanced crop management and breeding programs.
利用 RNA-seq 解密羽扇豆(Lupinus angustifolius)种子发育过程中的转录编程
羽扇豆(Lupinus spp.)种子因其蛋白质含量高(35-40%)而备受人类和动物的青睐。农作物种子的发育是影响种子命运和产量的关键因素,因此了解种子发育的分子机制至关重要。本研究对窄叶羽扇豆(NLL)种子发育阶段(开花后 3、6、9、12、15、18 和 21 天)的转录组进行了分析,以研究转录动态并确定控制种子发育的关键候选基因。从叶片、花和种子组织的九个样本中产生了约 3.57 亿个测序读数,从而鉴定出 34,769 个表达基因。分析表明,基因表达是动态的,早期以高代谢活动为特征,后期则以贮藏蛋白合成和营养库形成为重点。种子贮藏蛋白基因,包括杯蛋白类(α、β、γ 和 δ 凝集素)的表达模式差异显著。在后期(开花后 15-21 天),α 和 β 凝集素的表达量增加,这表明它们在谷粒充实和营养贮藏中发挥作用。与喹嗪类生物碱生物合成有关的基因,如赖氨酸/鸟氨酸脱羧酶和嘌呤渗透酶转运体 1,出现了晚期表达模式,表明生物碱的合成和转运在后期进行。许多在种子发育过程中发挥作用的成熟转录因子(TFs)(bHLH、AP2、MYB、ERF、C2H2、NAC、WRKY 和 C3H 锌指家族)都出现了差异表达,从而加强了我们研究结果的有效性。这些发现为了解羽扇豆种子发育的遗传和分子机制奠定了基础,有助于加强作物管理和育种计划。
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