籼稻背景下抑制不同ALK等位基因表达对稻米品质的影响
收稿日期: 2019-08-07
修回日期: 2019-09-13
网络出版日期: 2019-11-10
基金资助
国家自然科学基金资助项目(31825019);国家转基因重大专项(2016ZX08001006-005);江苏省科技计划资助项目(BE2018357)
Grain Quality as Affected by Down-regulation of Expression of Different ALK Alleles in indica Rice (Oryza sativa L.)
Received date: 2019-08-07
Revised date: 2019-09-13
Online published: 2019-11-10
目的 稻米糊化温度是影响稻米品质的重要指标,该性状受主效基因ALK/SSII-3调控,ALK基因具有多个复等位基因,本研究旨在通过RNAi技术明确籼稻亚种中两个不同ALK等位基因的效应。方法 以分别含有ALKc和ALKb等位基因的高糊化温度品种珍汕97B和低糊化温度品种龙特甫B为试验材料,使用RNAi技术构建ALK表达下调的转基因株系,通过对其稻米理化品质的测定来明确不同等位基因表达下调对稻米品质的影响。结果 对不同转基因水稻目的基因的表达分析显示本研究中转基因株系的ALK基因受到了不同程度的干扰。重点分析了不同RNAi株系稻米的糊化温度,结果表明珍汕97B的RNAi转基因稻米的糊化温度极显著降低,而在低糊化温度品种龙特甫B背景中下调表达ALK基因后对糊化温度的影响较小;转基因株系与未转化亲本相比,米粉的起始糊化温度都显著降低,表现为提前糊化;在珍汕97B背景下干扰系的峰值温度与未转化亲本相比极显著降低,而在龙特甫背景下米粉的峰值温度与未转化对照相比显著降低。对不同转基因系的理化品质分析表明,ALK下调表达植株稻米的表观直链淀粉含量显著增加,下调表达ALK后会引起米粉峰值黏度和崩解值的改变。高糊化温度品种珍汕97B干扰系与未转化对照相比胶稠度呈现极显著性差异,而低糊化温度品种龙特甫干扰系的胶稠度与未转化对照相比没有差异。结论 下调表达ALK等位基因对稻米理化品质产生显著影响,并且干扰不同等位基因的效应存在明显差异,即籼稻中的两个ALK等位基因的效应存在显著差异。
陈专专, 李先锋, 仲敏, 葛家奇, 范晓磊, 张昌泉, 刘巧泉 . 籼稻背景下抑制不同ALK等位基因表达对稻米品质的影响[J]. 中国水稻科学, 2019 , 33(6) : 513 -522 . DOI: 10.16819/j.1001-7216.2019. 9090
【Objective】 Gelatinization temperature (GT) is one of the key physicochemical properties in rice quality, which is mainly regulated by ALK (SSII-3) gene. In general, there are two ALK alleles among indica cultivars. To detect their functional differentiation in indica rice,【Method】 Zhenshan 97B (a high GT variety carrying ALKc allele) and Longtefu B (a low GT variety carrying ALKb allele), were used as receptors for the generation of transgenic rice with down-regulation of ALK expression by RNA interference (RNAi).【Result】 Down-regulation of ALK gene significantly decreased the GT of the transgenic lines. Due to the difference of original GT between the two receptors, the GT of transgenic rice lines derived from Zhenshan 97B (a high GT variety) decreased significantly, but it is slightly decreased in transgenic plants derived from Longtefu B (a low GT variety). The differential scanning calorimetry (DSC) results showed that the initial temperature of RNAi transgenic rice was significantly lower than the corresponding control and the transgenic lines were gelatinized in advance. The peak value of GT(Tp) in RNAi rice grains was significantly lower than that of the control under Zhenshan 97B background. However, Tp of RNAi rice grains under Longtefu B background was significantly lower than the control to a lesser extent. Also, down-regulation of ALK expression had a significant effect on rice physical-chemical characteristics. An increase of apparent amylose content in RNAi transgenic plants was detected due to the decreased expression of ALK gene. Besides, the pasting properties showed that down-regulation of ALK gene had obvious effects on peak viscosity and breakdown value, improving the taste of the transgenic rice. The gel consistency was significantly different among Zhenshan 97B RNAi lines and their parents, but no difference was found in Longtefu-derived transgenic lines.【Conclusion】 RNA interference to ALK allele expression had a significant effect on rice quality, especially the gelatinization characters. Down-regulated expression level of ALKc allele would cause larger variation of physical-chemical characteristics between transgenic rice and their parent than that of ALKb allele.
Key words： rice; ALK allele; RNA interference; gelatinization temperature; rice grain quality
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