中文摘要
缺硼和铝毒均是影响酸性土农业生产的重要问题,普遍认为施硼可以缓解铝毒,然而硼是否减少根尖铝累积尚存争议。我们前期的研究表明硼显著促进过渡区质外体碱性化,降低过渡区却促进伸长区铝的累积,其机理可能与生长素在过渡区的极性运输对质子泵的调控有关。利用LCSM、NMT、RT-PCR、FIA等技术,设计不同 pH(缓冲)环境,引入快速碱化因子RALF,研究过渡区质外体碱性化与铝毒的关系;利用FC、VA、突变体ralf1和fer4,研究过渡区碱性化与质子泵的关系;采用阻断生长素极性运输,利用生长素敏感元DII-VERNUS等,研究生长素运输对不同根区质膜H+-ATPase的调控;并利用囊泡运输抑制剂BFA和生长素感受突变体,研究硼和铝是否通过作用于生长素运输载体表达和循环及生长素信号感受而调节生长素极性运输。旨在更深入揭示硼缓解铝毒的内在机理。对于指导南方酸性土施肥,减轻土壤铝毒,具有重要实践意义。
英文摘要
Both boron deficiency and Al toxicity are the major factors affecting the agricultural productivity in acid soil. It is well known that B application alleviates Al toxicity, however, there are still debates on whether Al accumulation in roots can be reduced by B supply. Our previous research shows that B promotes the alkalinization and reduces Al accumulation in root transition zone while increase Al in elongation zone, which is hypothesized to be related to the regulation of plasma membrane H+-ATPase by auxin polar transport through root transition zone. Taking advantages of modern techniques such as LCSM, NMT, RT-PCR, FIA etc, the relationship between apoplast alkalinization and roles of B in alleviating Al toxicity will be studied under different pH situations with the adoption of rapid alkalinization factors (RALF) peptide. Relation between apoplast alkalinization and H+-ATPase will be carried out with the treatments of FC and VA, the stimulator and inhibitor of H+-ATPase, while adopting the mutants of RALF and its acceptor (ralf1 and fer4). The regulation of auxin transport on H+-ATPase will be studied by inhibiting the transporters of auxin efflux/influx by NPA/CHPAA or using mutants of the transporters (aux 1-7, eir 1-4). Auxin distribution will be detected in lines of auxin reporter and sensor, PIN2:GFP, DR5rev:GFP, DR5rev:GFP/pin2, DII-VENUS. Brefeldin (BFA), the inhibitor of exocytosis, will be used to study the mechanism of B and Al in regulation of auxin polar transport in the gene expression of PIN2 and AUX1, PIN2 recycling and auxin sensing. The mutants of auxin binding protein (abp1 mutants abp1-5 and its acceptor (rop6-1) will be used to confirm roles of auxin polar transport and auxin perception in regulation of H+-ATPase and apoplast alkalinization of root transition zone. The aims of the research are to disclose the mechanism of B in promoting apoplast alkalinization and alleviating Al toxicity to roots, thus to provide the understanding of the agricultural practice of B fertilization in alleviating Al toxicity besides supplying B, therefore to promote the productivity in acid soils.
