Background

Low temperature (LT) or cold stress is a major environmental stress that seriously affects plant growth and development, limiting crop productivity. Cold shock domain proteins (CSDPs), which are present in most living organism, are involved in RNA metabolisms influencing abiotic stress tolerance.

Objective

The aims of this study are to identify target gene for LT-tolerance, like CSDPs, characterize genetics, and develop molecular marker distinguishing LT-tolerance in cabbage (Brassica oleracea var. capitata).

Methods

Semi-quantitative RT-PCR or qRT-PCR was used in gene expression study. LT-tolerance was determined by electrolyte leakage and PCR with allelic specific primers.

Results

Allelic variation was found in BoCSDP5 coding sequence (CDs) between LT-tolerant (BN106 and BN553) and -susceptible inbred lines (BN107 and BN554). LT-tolerant inbred lines contained variant type of BoCSDP5 (named as BoCSDP5v) which encodes extra CCHC zinc finger domain at C-terminus. Association of LT-tolerance with BoCSDP5v was confirmed by electrolyte leakage and segregation using genetic population derived from BN553 and BN554 cross. Allelic variation in BoCSDP5 gene does not influence the rate of gene expression, but produces different proteins with different number of CCHC zinc finger domains. LT-tolerance marker designed on the basis of polymorphism between BoCSDP5 and BoCSDP5v was confirmed with samples used in previous B. oleracea CIRCADIAN CLOCK ASSOCIATED 1 (BoCCA1) marker validation.

Conclusions

LT-tolerant allele (BoCSDP5v) is dominant and independent of CBF pathway, and sufficient to generate molecular markers to identify LT-tolerant cabbage when it is used in combination with another marker, like BoCCA1-derived one. Production and analysis of overexpressing plants of BoCSDP1, BoCSDP3, BoCSDP5 and BoCSDP5v will be required for elucidating the function of CCHC zinc finger domains in LT-tolerance.

中文翻译：

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甘蓝型油菜冷激域蛋白5（BoCSDP5）富含甘氨酸的区域的自然变化与低温耐受性相关

背景

低温或低温胁迫是严重影响植物生长发育的主要环境胁迫，限制了作物的生产力。存在于大多数活生物体中的冷激域蛋白（CSDP）参与影响非生物胁迫耐受性的RNA代谢。

目的

这项研究的目的是确定目标基因LT-宽容，像CSDPs，鉴定遗传学，发展分子标记区分白菜LT-公差（甘蓝变种结球甘蓝）。

方法

半定量RT-PCR或qRT-PCR用于基因表达研究。通过电解质渗漏和使用等位基因特异性引物的PCR确定LT耐受性。

结果

在BoCSDP5编码序列（CD）中发现了耐LT耐受（BN106和BN553）和易感近交系（BN107和BN554）之间的等位基因变异。耐LT的近交系包含BoCSDP5的变体类型（命名为BoCSDP5v），该变体在C端编码额外的CCHC锌指结构域。通过使用来自BN553和BN554杂交的遗传种群进行电解质泄漏和分离，证实了LT耐受与BoCSDP5v的关联。BoCSDP5基因的等位基因变异不会影响基因表达的速率，但会产生具有不同数量CCCC锌指结构域的蛋白质。基于BoCSDP5与多态性之间的多态性设计的LT耐受标记BoCSDP5 v已通过先前的油菜花椰菜CIRCADIAN CLOCK ASSOCIATED 1（BoCCA1）标记验证中使用的样品进行了确认。

结论

耐LT的等位基因（BoCSDP5v）占主导地位，与CBF途径无关，当与另一种与BoCCA1衍生的标记结合使用时，足以产生分子标记来鉴定耐LT的甘蓝。为了阐明CCHC锌指结构域在LT耐受中的功能，需要生产和分析BoCSDP1，BoCSDP3，BoCSD P5和BoCSDP5v的过表达植物。