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Abstract
背景回顾：Hydrolyzable tannins (HTs), predominant polyphenols in oaks, are widely used in grape wine aging, feed additives, and human healthcare.
提出问题：However, the limited availability of a high-quality reference genome of oaks greatly hampered the recognition of the mechanism of HT biosynthesis.
主要研究及方法：Here, high-quality reference genomes of three Asian oak species (Quercus variabilis, Quercus aliena, and Quercus dentata) that have different HT contents were generated. Multi-omics studies were carried out to identify key genes regulating HT biosynthesis. In vitro enzyme activity assay was also conducted. Dual-luciferase and yeast one-hybrid assays were used to reveal the transcriptional regulation.
结果1-水解单宁生化标志物及其关键合酶：Our results revealed that β-glucogallin was a biochemical marker for HT production in the cupules of the three Asian oaks. UGT84A13 was confirmed as the key enzyme for β-glucogallin biosynthesis.
结果2-三个亚洲栎中水解单宁合成的转录调控分化：The differential expression of UGT84A13, rather than enzyme activity, was the main reason for different β-glucogallin and HT accumulation. Notably, sequence variations in UGT84A13 promoters led to different trans-activating activities of WRKY32/59, explaining the different expression patterns of UGT84A13 among the three species.
结论：Our findings provide three high-quality new reference genomes for oak trees and give new insights into different transcriptional regulation for understanding β-glucogallin and HT biosynthesis in closely related oak species.
摘 要
水解单宁是橡树中主要的多酚类物质，被广泛用于葡萄酒陈酿、饲料添加剂和人类医疗保健。然而，由于缺乏橡树高质量参考基因组，极大地阻碍了对水解单宁生物合成机制的认识。本文中，作者组装获得了栓皮栎、槲栎、槲树三个具有不用水解单宁的亚洲栎的高质量基因组。作者通过多组学分析鉴定了调节水解单宁生物合成的关键基因，并通过体外酶活试验进行了验证，以及通过双荧光素酶和酵母单杂交试验进行转录调控研究。结果表明，β-没食子酰葡萄糖是三种亚洲栎壳斗中水解单宁生成的生化标志物。UGT84A13被证实为β-没食子酰葡萄糖生物合成的关键酶。相比于UGT84A13的酶活，其编码基因的差异表达更有可能是导致β-没食子酰葡萄糖和水解单宁积累不同的主要原因。值得注意的是，UGT84A13启动子序列变异是导致WRKY32/59介导的反式激活差异的关键原因，这解释了UGT84A13在三个亚洲栎中的不同表达模式。本文的研究结果为橡树提供了三个高质量的新参考基因组，并为理解近缘橡树物种中β-没食子酰葡萄糖和水解单宁生物合成的不同转录调控提供了新的见解。
北京林业大学李国雷教授为本文通讯作者，北京林业大学青年教师杨钦淞、博士生郦金今、中国农业大学博士后王妍为论文的共同第一作者。该研究得到了国家重点研发计划（2021YFD2200302）、国家自然科学基金（32201544）以及中国博士后科学基金（2021M700451）的联合资助。
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