Proteaceae, a largely southern hemisphere family consisting of 80 genera distributed in Australia and southern Africa as its centres of greatest diversity, also extends well in northern and southern America. Under this family, Grevillea robusta is a fast-growing species got popularity in farm and avenue plantations. Despite the ecological and economic importance, the species has not yet been investigated for its genetic improvement and genome-based studies. Only a few molecular markers are available for the species or its close relatives, which hinders genomic and population genetics studies. Genetic markers have been intensively applied for the main strategies in breeding programs, especially for the economically important traits. Hence, it is of utmost priority to develop genomic database resources and species-specific markers for studying quantitative genetics in G. robusta. Given this, the present study aimed to develop de novo genome sequencing, robust microsatellites markers, sequence annotation and their validation in different stands of G. robusta in northern India. Library preparation and sequencing were carried out using Illumina paired-end sequencing technology. Approximately, ten gigabases (Gb) sequence data with 70.87 million raw reads assembled into 425,923 contigs (read mapped to 76.48%) comprising 455 Mb genome size (23 × coverage) generated through genome skimming approach. In total, 9421 simple sequence repeat (SSR) primer pairs were successfully designed from 13,335 microsatellite repeats. Afterward, a subset of 161 primer pairs was randomly selected, synthesized and validated. All the tested primers showed successful amplification but only 13 showed polymorphisms. The polymorphic SSRs were further used to estimate the measures of genetic diversity in 12 genotypes each from the states of Punjab, Haryana, Himachal Pradesh and Uttarakhand. Importantly, the average number of alleles (N_a), observed heterozygosity (H_o), expected heterozygosity (H_e), and the polymorphism information content (PIC) were recorded as 2.69, 0.356, 0.557 and 0.388, respectively. The availability of sequence information and newly developed SSR markers could potentially be used in various genetic analyses and improvements through molecular breeding strategies for G. robusta.

山龙眼科主要是南半球的一个科，由 80 个属组成，分布在澳大利亚和南部非洲，是其多样性最丰富的中心，在美洲北部和南部也有广泛分布。在这个家族中，银桦是一种快速生长的物种，在农场和街道种植园中很受欢迎。尽管具有生态和经济重要性，但尚未对该物种进行遗传改良和基于基因组的研究。该物种或其近亲只有少数分子标记可用，这阻碍了基因组和群体遗传学研究。遗传标记已被广泛应用于育种计划的主要策略，特别是对于经济上重要的性状。因此，开发基因组数据库资源和物种特异性标记用于研究鲁布斯塔的数量遗传学是当务之急。鉴于此，本研究旨在开发从头基因组测序、强大的微卫星标记、序列注释及其在印度北部不同地区的G.Robusta中的验证。使用Illumina双端测序技术进行文库制备和测序。大约 10 GB 序列数据（包含 7087 万条原始读数）组装成 425,923 个重叠群（读数映射到 76.48%），其中包含通过基因组略读方法生成的 455 Mb 基因组大小（23 × 覆盖率）。总共，从 13,335 个微卫星重复中成功设计了 9421 个简单序列重复 (SSR) 引物对。随后，随机选择、合成并验证了 161 个引物对的子集。所有测试的引物均显示成功扩增，但只有 13 个显示多态性。 多态性 SSR 进一步用于估计来自旁遮普邦、哈里亚纳邦、喜马偕尔邦和北阿坎德邦的 12 种基因型的遗传多样性测量。重要的是，平均等位基因数 (N _a )、观察到的杂合度 (H _o )、预期杂合度 (H _e ) 和多态性信息内容 (PIC) 分别记录为 2.69、0.356、0.557 和 0.388。序列信息和新开发的 SSR 标记的可用性有可能用于各种遗传分析，并通过粗麻布的分子育种策略进行改进。