Identification of Dehydrin-Like Proteins Responsive to Chilling in Floral Buds of Blueberry (Vaccinium, section Cyanococcus)
Identification of Dehydrin-Like Proteins Responsive to Chilling in Floral Buds of Blueberry (Vaccinium, section Cyanococcus)
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DOI：
10.1104/pp.104.4.1439
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发表时间：
1994-04
期刊：
影响因子：
--
通讯作者：
M. Muthalif;L. Rowland
中科院分区：
文献类型：
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作者：
M. Muthalif;L. Rowland
三个主要的多肽的65，60，和14 kD的水平增加响应于低温单位积累在花芽的木本多年生植物，蓝莓（越橘，节蓝球藻）。在低温处理后300 h内，这些多肽的含量增加最明显，随着芽萌发的开始，这些多肽的含量下降到低温前的水平。以伞房越桔和灰叶越桔的休眠芽为材料，研究了不同低温处理后休眠芽的抗寒性。这些水平与寒冷响应多肽的水平一致。与一年生植物中的其他冷诱导蛋白一样，叶片中冷诱导多肽的水平也响应于冷处理而增加;冷冻诱导的多肽是热稳定的，在95[deg]C温育15分钟后抗聚集。通过首先通过等电点（pl）然后通过分子量分级芽蛋白，发现65-和60-kD多肽的pI值为7.5 - 8.0，14-kD多肽的pI值判断为8.5。纯化的65-和60-kD的多肽，然后用内切蛋白酶Lys-C消化和选定的片段的测序，揭示了65-和60-kD的多肽之间的氨基酸组成的相似性，和Ephinins。事实上，抗血清富含赖氨酸的共识序列EKKGIMDKIKEKLPG的蓝莓蛋白交叉反应的所有三个主要的冷冻响应多肽的蓝莓，确定这些作为蓝莓蛋白或类蓝莓蛋白。
The level of three major polypeptides of 65, 60, and 14 kD increased in response to chilling unit accumulation in floral buds of a woody perennial, blueberry (Vaccinium, section Cyanococcus). The level of the polypeptides increased most dramatically within 300 h of chilling and decreased to the prechilling level with the initiation of budbreak. Cold-hardiness levels were assessed for dormant buds of Vaccinium corymbosum and Vaccinium ashei after different chilling treatments until the resumption of growth. These levels coincided with the level of the chilling-responsive polypeptides. Like some other previously described cold-induced proteins in annual plants, the level of the chilling-induced polypeptides also increased in leaves in response to cold treatment; the chilling-induced polypeptides were heat stable, resisting aggregation after incubation at 95[deg]C for 15 min. By fractionating bud proteins first by isoelectric point (pl) and then by molecular mass, the pl values of the 65- and 60-kD polypeptides were found to be 7.5 to 8.0 and the pl value of the 14-kD polypeptide was judged to be 8.5. Purification of the 65- and 60-kD polypeptides, followed by digestion with endoproteinase Lys-C and sequencing of selected fragments, revealed similarities in amino acid composition between the 65- and 60-kD polypeptides and dehydrins. Indeed, antiserum to the lysine-rich consensus sequence EKKGIMDKIKEKLPG of dehydrin proteins cross-reacted to all three of the major chilling-responsive polypeptides of blueberry, identifying these as dehydrins or dehydrin-like proteins.