干旱条件下外源蔗糖对高表达玉米C4pepc水稻种子萌发的影响
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国家自然科学基金(31571585)


Physiological Mechanism on Seed Germination in C4 Phosphoenolpyruvate Carboxylase Transgenic Rice (Oryza sativa) by Exogenous Sucrose under Drought Conditions
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    摘要:

    为揭示外源蔗糖参与干旱胁迫下高表达转玉米C4 型磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase, PEPC)基因(C4pepc)水稻(简称:PC)种子萌发的生理机制,该研究以 PC及其未转基因野生型受体‘Kitaake’(简称:WT)的种子为材料,研究外施不同浓度蔗糖联合模拟干旱(10% PEG6000)处理下,其种子发芽参数、总可溶性糖及可溶性蛋白含量、蔗糖非发酵1 (sucrose nonfermenting1, SNF1)相关蛋白激酶(SNF1related protein kinase 1s, SnRK1s)基因以及PEPC基因表达等参数的变化。结果表明:(1)PEG6000模拟干旱处理均显著抑制两材料发芽,但明显促进胚根的生长;外施蔗糖则呈现浓度效应,高浓度蔗糖(>150 mmol·L-1)进一步加剧了干旱对发芽的抑制效应,而低浓度(<30 mmol·L-1)则可缓解干旱的抑制,但与WT(<30 mmol·L-1)相比,促进PC水稻萌发的外施蔗糖浓度(<6 mmol·L-1)更低,且各处理的发芽表现与其α淀粉酶活性的动态表现一致。(2)与WT相比,外施3 mmol·L-1蔗糖联合干旱处理下,显著提高了PC种子的发芽率,且伴随PC内源蔗糖含量、总可溶糖和可溶性蛋白含量显著增加;且外施3 mmol·L-1蔗糖使PC中内源C3pepc基因表达下调,而外源导入C4pepc基因表达显著增加。(3)与WT相比,干旱处理下外施3 mmol·L-1蔗糖,PC的糖信号相关基因SnRKs亚家族基因(包括SnRK1s:OsK1a OsK24 OsK35和SnRK2s:SAPK6)的表达也显著增加。研究发现,外施低浓度蔗糖通过上调PC水稻种子中可溶性糖和可溶性蛋白含量,增强SnRK1s亚家族基因和外源C4pepc基因的表达,提高了α淀粉酶活性,从而缓解了干旱胁迫对PC种子萌发的抑制。

    Abstract:

    In order to investigate the intrinsic mechanism of sucrose participated in drought tolerance in plants, the study used the phosphoenolpyruvate carboxylase (C4pepc) transgenic rice (Hereinafter referred to as PC) and untransformed wildtype “Kitaake” (Hereinafter referred to as WT) rice lines. PC and WT were treated with different concentrations of sucrose during seed germination under simulated drought treatment (10% PEG6000). The performance of seed germination, total soluble sugar and sugar components contents, total soluble protein content, transcript levels of genes of sucrose nonfermenting1(SNF1)related protein kinases (SnRKs) and transcript levels of PEPC genes in rice lines were measured. The results showed as following: (1) external application of sucrose solution showed the similar effect with the seed germination of rice, but significantly promoted the seed germination with low concentration of sucrose solution companying with drought treatment. External application of sucrose solution showed a concentrationdependent effect on seed germination of two rice lines. High sucrose concentration (>150 mmol·L-1) further inhibited drought effect, while low concentration (<30 mmol·L-1) alleviated drought inhibition. However, the sensitive concentration of sucrose with the best effect on seed germination was different in two lines under drought treatment, PC (<6 mmol·L-1) is much lower than WT (<30 mmol·L-1). And this performance is consistent with the dynamic changes of the activity of αamylase in two lines as well. (2) Compared with WT, 3 mmol·L-1 sucrose combined with drought treatment significantly promoted the performance of seed germination in PC with a significant increase of endogenous sucrose content, total soluble sugar content and soluble protein content. Furthermore, the treatment of 3 mmol·L-1 sucrose with drought significantly decreased the transcript level of C3pepc, but significantly increased C4pepc in the PC, respectively (P<0.05); (3) Some sugar signaling related genes such as SnRKs family genes including SnRK1s (OsK1a OsK24 OsK35) and SnRK2s (SAPK6) in PC rice also significantly increased their transcript levels after the same treatment as above. The results showed that PC can increase the contents of soluble sugar and soluble protein, regulate the expression of SnRKs subfamily genes and exogenous C4pepc by increasing low sucrose, improve the activity of αamylase, and therefore alleviate the inhibitory effect on the seed germination under drought treatment.

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张晓敬,李 霞,吴博晗,等.干旱条件下外源蔗糖对高表达玉米C4pepc水稻种子萌发的影响[J].西北植物学报,2020,40(11):1888-1899

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  • 在线发布日期: 2020-12-11
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