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以普通花生品种花育22为母本、高油酸花生品种开农176为父本杂交得到F1杂种,筛选油酸含量高于60%且同时含有FAD2a和FAD2b位点的F1为杂交父本,以花育22为轮回亲本(母本)连续回交得到BC1F1~BC4F1代回交种。利用近红外光谱仪测定F1及BC1F1~BC4F1籽粒的油酸、亚油酸含量,选择油酸含量大于60%的种子,用刀片切取种子小部分子叶提取DNA,以F0.7/R3为引物进行PCR扩增及测序,根据测序峰图差异表现筛选出同时含有FAD2a和FAD2b位点的种子作为下一代回交的父本。切去部分子叶的种子切口用石蜡封闭,播种前浸泡于40℃温水中催芽,对12 h后未露白的种子用100 mg L–1乙烯利浸泡4 h后再转入40℃温水浸泡至24 h,发芽率可达到98%。2013年春季开始杂交,2016年春在青岛播种BC4F2代种子,取幼苗期幼叶鉴定基因型,筛选出基因型为aabb的单株,收获时选留农艺性状类似于花育22的优良单株,再利用近红外光谱仪测定所选单株油酸含量,获得油酸含量在70%以上、油酸亚油酸比值大于7.0的单株24个。这些单株与花育22相比,农艺性状基本相同,称为改良花育22高油酸花生新品系。
F1 hybrids were obtained from Huayu 22 with the same peanut variety Huayu22 and F1 hybrid with high oleic peanut variety Kai Nong 176 as the male parent. The F1 hybrids containing more than 60% oleic acid and containing FAD2a and FAD2b sites were selected as hybrid parents. Huayu 22 as the reincarnated parent (female) continuous backcross BC1F1 BC4F1 back to the exchange. The oleic acid and linoleic acid content of F1 and BC1F1 ~ BC4F1 seeds were determined by near infrared spectroscopy, and the seeds with oleic acid content greater than 60% were selected. The small cotyledons of seeds were extracted with a razor blade to extract DNA. F0.7 / R3 was used as primer Amplified and sequenced. The seeds containing both FAD2a and FAD2b sites were screened out as the male-backcross of the next generation based on the differential expression of sequencing peak maps. Sections of cut cotyledons were cut in paraffin and sown in warm water at 40 ° C prior to sowing. After 12 h of exposure, untreated seeds were soaked in 100 mg L-1 ethephon for 4 h and then immersed in warm water at 40 ° C until 24 h, the germination rate can reach 98%. The hybridization began in the spring of 2013 and the BC4F2 generation seed was sown in Qingdao in the spring of 2016. The genotypes of the young leaves of the seedling were identified and the individual plants with the genotype aabb were screened. The elite agronomic traits similar to Huayu22 were selected at harvest. Then the content of oleic acid in the selected plant was determined by near infrared spectroscopy to obtain 24 plants with oleic acid content above 70% and oleic acid to linoleic acid ratio over 7.0. Compared with Huayu 22, the agronomic traits of these plants are basically the same, which is called the new line of improved oleate peanut of Huayu 22.