普通小麦中国春-百萨偃麦草异染色体系的分子标记分析

Molecular Marker Analysis on Common Wheat Landrace Chinese Spring Alien Chromosome Lines Derived from Thinopyrum bessarabicum Lve

综合利用HMW-Glu亚基、STS、SSR和RFLP等分子标记对普通小麦中国春、百萨偃麦草、中国春-百萨偃麦草双二倍体和11个中国春-百萨偃麦草异染色体系进行了分析。结果表明,14对SSR、10对STS引物和6个RFLP标记可以特异追踪百萨偃麦草染色质。C7-17及其后代株系C7-17-2等编码百萨偃麦草特异HMW-Glu亚基,添加染色体涉及与小麦第1部分同源群染色体部分同源的1J;1对STS、3对SSR和1个RFLP探针可以特异追踪二体附加系CH05中的百萨偃麦草染色体,并揭示最初根据分带核型确定的J3与小麦第2部分同源群染色体具有较高的部分同源性;2对STS、1个RFLP探针和1对SSR可以追踪CH09的外源染色体,并揭示最初确定的J7与小麦第3部分同源群染色体具有较高的部分同源性;1对STS和1个RFLP探针在CH03、CH04和CH34中具有相同的多态,3个附加系可能添加了相同染色体,最初确定的J1、J2和J?与小麦第7部分同源群染色体具有较高的部分同源性;3对SSR引物可以特异追踪CH12中附加的大片段易位染色体和CH11中的小片段易位染色体,推测易位可能涉及同一条百萨偃麦草染色体。发现13个标记(5个STS、3个RFLP探针和5个SSR)可以追踪未涉及到的4J和5J等染色体。

Thinopyrum Mssarabicum Loeve and Chinese Spring-Thinopyrtun Mssarabicum amphiploid （the amphiploid hereafter） are reported with high salinity tolerance. Screen of molecular markers tagging different chromosemes of Thinopyrum bessarabicum would facilitate transfer of these genes or chromosomes into common wheat. Some specific proteins or enzymes, AFLP or RAID markers have been described by different authors. However, use of SSR, STS, and RFLP markers identifying Thinopyrum bessarabicum chromosomes is seldom reported. To dig out the beneficial genes of this species, the amphiploid was introduced from Dr. Mujeeb-Kazi, CIMMYT, and several additions, and translocations have been independently developed and reported by Cytogentics Institute, Nanjing Agricultural University, China. Using SSR, STS, and RFLP markers of wheat and barley etc, 7 disomic additions, 3 alien lines including translocations and additions and 1 multiple additions were characterized in this study. The results revealed that 14 of 219 wheat SSRs, 8 of 126 barley STS and 2 wheat STS, 6 RFLP probes could be used to distinguish the ehromosomes of Thinopyrum bessarabicum added in wheat. Of the alien chromosome lines, C7-17 and C7-17-2 were verified involving chromosome 1J encoding specific HMW- Glu subunit of Thinopyrmn bessarabicum. Two STS markers MWG2303 （2HL） and NAU/Xibaol5 （6S）, one RFLP probe BCD135 （2BE） and4 SSR markers （Cfd168 （2A）, Xgwm526 （2B）, Xgwm311 （2D）, and Xgdm93 （4B）） could be used to tag the chromosomes added in CH05, while 6L probe PSR154 and one wheat 2AL-end STS KsuH9 failed to produce polymorphism indicating rearrangements occurred in this chromosome, therefore, the preliminary designed J3 based on the C-banded karyotype of Thinopyrmn bessarabicum added in this line was deduced with closer relationship with group 2 chromosomes of wheat. Two STSs from 3HL（BCD269, MWG974）, one probe PSRll6 from wheat 3L and one SSR Xgwmll4（3DS） could be used to tag the chromosomes added in CH09, the preliminary designed J7 was then deduced with closer relationship with group 3 chromosomes of wheat. One STS MWG808（7H）, one probe mapped in the middle part of 4L（PSR163） and one SSR Xgdml31（3B） could be used to tag the chromosomes added in CH03,CH04 and CH34, while two other probes mapped in the distal part of 4L （WG114） and 4S （CD0669） without RFLPs, indicating these three additions might contain the same alien chromosomes, the preliminary designed J1, J2, and J？ in these lines respectively were deduced with closer relationship with group 7 chromosomes of wheat, rearrangements with group 4 chromosomes also involved. Three SSR markers Xgdm35 （2D）, Xgwm515 （2A, 2D）, and Xgwm614 （2A, 2B, and 2D） could be used to tag the large segmental translocation chromosomes added in CH12, and small segmental translocation chromosomes contained in CHll which also included another pair of alien chromosomes similar to that of CH05, this result indicated that these translocations with different segments might be derived from the same chromosome of Th/nopyrum bessarabicum. 13 markers （5 barley STSs, 3 RFLP probes and 5 wheat SSRs） could be used to trace the chromosomes not involved in the present lines.