SRAP Marker Associated with Fusarium-wilt Resistance Gene in Sweet Potato
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摘要: 以高抗甘薯蔓割病品种金山57和高感蔓割病甘薯品种新种花为亲本进行杂交，获得F1分离群体，对F1群体进行蔓割病抗性鉴定，从中选出7个高抗和7个高感蔓割病株系构建抗、感池。以抗、感池基因组DNA为模板，用276对SRAP引物组合对其扩增，从中筛选出98对具有多态性的引物组合，再以抗感亲本加抗感池基因组DNA为模板筛选，从98份SRAP引物组合中筛选出33对多态性较好的引物组合。后通过用一组小群体（6个高抗株系和6个高感株系）进一步筛选，最终获得一对与甘薯抗蔓割病连锁的SRAP引物组合a12b8，经过抗感亲本及F1代的进一步验证，认为该标记与甘薯抗蔓割病基因相关。Abstract: Fusarium wilt (F.W) disease-resistant molecular markers can be used to screen disease-resistant lines in early planting stage of sweet potato, which can also greatly improve the efficiency of breeding for sweet potato disease-resistance.The F1 progenies were obtained from the cross of sweet potato c.v. Jinshan57, which is higly resistant to Fusarium wilt (F.W), and c.v. Xinzhonghua, which is highly susceptible to F.W. After disease-resistant identification, 7 highly disease-resistant and 7 highly disease-susceptible lines were selected from F1 progenies and were used to establish the resistant and susceptible DNA pools. The genomic DNA of the resistant and susceptible pools were PCR-amplified respectively with 276 pairs of SRAP primers, and 98 pairs of primer generated polymorphic bands between the disease-resistant and susceptible pools. Through further screening using a group of small population and their parents, we got a SRAP marker named a12b8, which would be linked to the resistance of F.W. in sweet potato. This marker would be used for identifying the resistance to F.W in the early growth stage of sweet potato and also would be utilized for improving the efficiency of F.W disease-resistant breeding by molecular marker assisted selection for sweet potato.
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图 1 用抗、感池筛选SRAP引物
注：1为抗病池；2为感病池；A~V为不同SRAP引物组合; M为Marker。
Figure 1. Screening of SRAP primers in FW-resistant and FW-susceptible pools
图 2 用抗、感亲本和抗、感池筛选SRAP引物
注：1为抗病池；2为感病池；a为抗病亲本；b为感病亲本；A~N为不同SRAP引物组合; M为Marker。
Figure 2. Screening of SRAP primers in parents and FW-resistant and FW-susceptible pools
图 3 小群体DNA对SRAP引物的进一步筛选
注：A~H为不同的引物组合；M为Marker；1~6为高抗株系；7~12为高感株系。
Figure 3. Secondary screening of SRAP primers with small group DNA
图 4 引物组合a12b8对F1群体部分单株的SRAP扩增结果
注：M为Marker; a为高抗亲本；b为高感亲本；1~11为高抗株系；12~22为高感株系。
Figure 4. SRAP-PCR amplification by a12b8 marker in partial F1 population
图 5 引物a12b8对F1群体其他株系扩增结果
注：M为Marker; A为抗病株系；B为感病株系。
Figure 5. SRAP-PCR amplification by a12b8 marker in remaining F1 population
表 1 SRAP分析所用的引物
Table 1. Primers used in SRAP analysis
引物 序号 序列 上游引物 a1 TGAGTCCAAACCGGCAG a2 TGAGTCCAAACCGGTAA a3 TGAGTCCAAACCGGTTT a4 TGAGTCCAAACCGGTTG a5 TGAGTCCAAACCGGAGT a6 TGAGTCCAAACCGGTTC a7 TGAGTCCAAACCGGTCT a8 TGAGTCCAAACCGGAGG a9 TGAGTCCAAACCGGAGC a10 TGAGTCCAAACCGGCAC a11 TGAGTCCAAACCGGACC a12 TGAGTCCAAACCGGATT 下游引物 b1 GACTGCGTACGAATTACA b2 GACTGCGTACGAATTCTT b3 GACTGCGTACGAATTCAG b4 GACTGCGTACGAATTACC b5 GACTGCGTACGAATTATC b6 GACTGCGTACGAATTGAA b7 GACTGCGTACGAATTAAG b8 GACTGCGTACGAATTACA b9 GACTGCGTACGAATTCTA b10 GACTGCGTACGAATTCAC b11 GACTGCGTACGAATTGAT b12 GACTGCGTACGAATTATA b13 GACTGCGTACGAATTCTC b14 GACTGCGTACGAATTGAA b15 GACTGCGTACGAATTCAA b16 GACTGCGTACGAATTCTA b17 GACTGCGTACGAATTGCA b18 GACTGCGTACGAATTCTG b19 GACTGCGTACGAATTCGC b20 GACTGCGTACGAATTGAG b21 GACTGCGTACGAATTCCC b22 GACTGCGTACGAATTGCG b23 GACTGCGTACGAATTCCT 
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