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赤霉素相关顺式基因改变杨树的生长、株型和木材特性。

Gibberellin-associated cisgenes modify growth, stature and wood properties in Populus.

机构信息

Department of Forest Ecosystems and Society, Oregon State University, Corvallis, OR, USA.

出版信息

Plant Biotechnol J. 2011 Feb;9(2):162-78. doi: 10.1111/j.1467-7652.2010.00537.x.

Abstract

We studied the effects on plant growth from insertion of five cisgenes that encode proteins involved in gibberellin metabolism or signalling. Intact genomic copies of PtGA20ox7, PtGA2ox2,Pt RGL1_1, PtRGL1_2 and PtGAI1 genes from the genome-sequenced Populus trichocarpa clone Nisqually-1 were transformed into Populus tremula × alba (clone INRA 717-1B4), and growth, morphology and xylem cell size characterized in the greenhouse. Each cisgene encompassed 1-2 kb of 5' and 1 kb of 3' flanking DNA, as well as all native exons and introns. Large numbers of independent insertion events per cisgene (19-38), including empty vector controls, were studied. Three of the cisgenic modifications had significant effects on plant growth rate, morphology or wood properties. The PtGA20ox7 cisgene increased rate of shoot regeneration in vitro, accelerated early growth, and variation in growth rate was correlated with PtGA20ox7 gene expression. PtRGL1_1 and PtGA2ox2 caused reduced growth, while PtRGL1_2 gave rise to plants that grew normally but had significantly longer xylem fibres. RT-PCR studies suggested that the lack of growth inhibition observed in PtRGL1_2 cisgenic plants was a result of co-suppression. PtGAI1 slowed regeneration rate and both PtGAI1 and PtGA20ox7 gave rise to increased variance among events for early diameter and volume index, respectively. Our work suggests that cisgenic insertion of additional copies of native genes involved in growth regulation may provide tools to help modify plant architecture, expand the genetic variance in plant architecture available to breeders and accelerate transfer of alleles between difficult-to-cross species.

摘要

我们研究了插入 5 个顺式基因对植物生长的影响,这些基因编码参与赤霉素代谢或信号转导的蛋白质。从已测序的毛果杨克隆 Nisqually-1 基因组中插入 PtGA20ox7、PtGA2ox2、PtRGL1_1、PtRGL1_2 和 PtGAI1 基因的完整基因组拷贝,转化到欧洲山杨×白杨(克隆 INRA 717-1B4)中,并在温室中对生长、形态和木质部细胞大小进行了特征描述。每个顺式基因包含 1-2 kb 的 5'侧翼 DNA 和 1 kb 的 3'侧翼 DNA,以及所有天然的外显子和内含子。每个顺式基因都研究了大量的独立插入事件(19-38 个),包括空载体对照。其中 3 个顺式基因修饰对植物生长速度、形态或木材特性有显著影响。PtGA20ox7 顺式基因增加了体外芽再生的速度,加速了早期生长,生长速度的变化与 PtGA20ox7 基因的表达相关。PtRGL1_1 和 PtGA2ox2 导致生长速度降低,而 PtRGL1_2 导致植物正常生长,但木质部纤维显著变长。RT-PCR 研究表明,PtRGL1_2 顺式基因植物中观察到的生长抑制缺失是共抑制的结果。PtGAI1 降低了再生速度,而 PtGAI1 和 PtGA20ox7 分别导致早期直径和体积指数的事件间方差增加。我们的工作表明,顺式基因插入额外的天然生长调节基因拷贝可能为改良植物结构提供工具,扩大可供育种者使用的植物结构遗传变异,并加速难以杂交物种之间的等位基因转移。

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