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作者	单位
李婷婷，姚有华，安立昆，白羿雄，杨雪，姚晓华，吴昆仑	（1.青海大学农林科学院，青海西宁 810016 2.青藏高原种质资源研究与利用实验室/青海省青稞遗传育种重点实验室/国家麦类改良中心青海青稞分中心，青海西宁 810016）

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中文摘要:

为探索青稞（Hordeum vulgare L. var. nudum Hook. f）WAK类基因在青稞抗条纹病中的作用，以抗条纹病青稞品种昆仑14号和感病品种Z1141为材料，从叶片中克隆了HvnWAK基因。昆仑14号HvnWAK基因开放阅读框（ORF）为2 964 bp，Z1141的ORF为3 048 bp，分别编码988个氨基酸和1 016个氨基酸，两者氨基酸序列一致性为97.14%。启动子区域预测表明，该区段包含脱落酸、茉莉酸、赤霉素等与逆境胁迫相关的多个顺式作用元件。蛋白质序列分析表明，HvnWAK为亲水性的不稳定酸性蛋白，具有典型的细胞壁相关激酶结构特征，如细胞壁受体结构（GUB_WAK_bind）、胞内激酶结构域（intracellular kinase structural domain）和跨膜域、表皮生长因子结构域（epidermal growth factor，EGF），属于WAK家族。同源比对与系统进化分析表明，昆仑14号的HvnWAK蛋白与大麦、山羊草、硬直黑麦草的WAK蛋白序列相似性分别为97.24%、92.17%、80.80%;Z1141的HvnWAK蛋白与大麦、山羊草、硬直黑麦草等的WAK蛋白序列相似性分别为100%、92.17%、80.80%。青稞HvnWAK蛋白与大麦HvWAK蛋白的亲缘关系最近，与玉米和水稻的亲缘关系较远。蛋白互作预测结果表明，与WAK蛋白存在互作关系的有抗病蛋白同源物、解旋酶家族蛋白、苏氨酸蛋白激酶等。实时荧光定量（qRT-PCR）分析表明，该基因在感病后迅速表达，且在抗病材料昆仑14号中的表达峰值显著高于Z1141（P＜0.01），且高峰时期（8～9周）晚于在Z1141中的表达高峰时期（7～8周）。由此推测，HvnWAK基因在青稞抗条纹病的后期发挥重要的调控作用。

英文摘要:

To explore the role of WAK-like genes in barley (Hordeum vulgare L. var. nudum Hook. f) in leaf stripe resistance, with a leaf stripe resistant barley variety Kunlun 14 and a susceptible variety Z1141 as materials, HvnWAK genes were cloned from leaves of the two hulless barley varieties. The open reading frame (ORF) of the HvnWAK gene of Kunlun 14 was 2 964 bp and the ORF of Z1141 was 3 048 bp, encoding 988 amino acids and 1 016 amino acids, respectively, with an amino acid sequence identity of 97.14%. The promoter region prediction indicated that this region contained multiple cis-acting elements of abscisic acid, jasmonic acid, and gibberellin associated with adversity stress, et al. Protein sequence analysis showed that HvnWAK was a hydrophilic unstable acidic protein with typical cell wall-associated kinases, such as cell wall receptor structure(GUB_WAK_bind), intracellular kinase structural domain(intracellular kinase structural domain), transmembrane domain, and epidermal growth factor(EGF) structural domain, which belonged to the WAK family. Homology comparison and phylogenetic analysis showed that the HvnWAK protein of Kunlun 14 had 97.24%, 92.17%, and 80.80% sequence similarity with the WAK proteins of barley, Aegilops tauschii, and Lolium rigidum, respectively; Meanwhile the HvnWAK protein of Z1141 had 100%, 92.17%, and 80.80% sequence similarity with the WAK proteins of barley, Aegilops tauschii, and Lolium rigidum, respectively. The HvnWAK protein was most closely related to the barley HvWAK protein and more distantly related to maize and rice. The predicted results of protein interactions showed that interactions with WAK proteins existed with disease resistance protein homologs, decapping enzyme family proteins, and threonine protein kinases. Quantitative real-time fluorescence(qRT-PCR) analysis showed that the gene was rapidly expressed after susceptibility to disease, and the peak in the disease-resistant material Kunlun 14 was significantly higher than that in Z1141(P＜0.01), and the peak period (8—9 weeks) was later than that in Z1141 (7—8 weeks). It is hypothesized that the HvnWAK gene plays an important regulatory role in the later stages of leaf stripe resistance in hulless barley.

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