Gupta Sudhir Kumar, Charpe Ashwini, Prabhu Kumble Vinod, Haque Qazi Mohammad Rizwanul
National Phytotron Facility, Indian Agricultural Research Institute, New Delhi 110012, India.
Theor Appl Genet. 2006 Oct;113(6):1027-36. doi: 10.1007/s00122-006-0362-7. Epub 2006 Aug 1.
A leaf rust resistance gene Lr19 on the chromosome 7DL of wheat derived from Agropyron elongatum was tagged with random amplified polymorphic DNA (RAPD) and microsatellite markers. The F(2) population of 340 plants derived from a cross between the leaf rust resistant near-isogenic line (NIL) of Thatcher (Tc + Lr19) and leaf rust susceptible line Agra Local that segregated for dominant monogenic leaf rust resistance was utilized for generating the mapping population. The molecular markers were mapped in the F(2) derived F(3) homozygous population of 140 seedlings. Sixteen RAPD markers were identified as linked to the alien gene Lr19 among which eight were in a coupling phase linkage. Twelve RAPD markers co-segregated with Lr19 locus. Nine microsatellite markers located on the long arm of chromosome 7D were also mapped as linked to the gene Lr19, including 7 markers which co-segregated with Lr19 locus, thus generating a saturated region carrying 25 molecular markers linked to the gene Lr19 within 10.2 +/- 0.062 cM on either side of the locus. Two RAPD markers S265(512) and S253(737) which flanked the locus Lr19 were converted to sequence characterized amplified region markers SCS265(512) and SCS253(736), respectively. The marker SCS265(512) was linked with Lr19 in a coupling phase and the marker SCS253(736) was linked in a repulsion phase, which when used together mimicked one co-dominant marker capable of distinguishing the heterozygous resistant seedlings from the homozygous resistant. The molecular markers were validated on NILs mostly in Thatcher background isogenic for 44 different Lr genes belonging to both native and alien origin. The validation for polymorphism in common leaf rust susceptible cultivars also confirmed the utility of these tightly linked markers to the gene Lr19 in marker-assisted selection.
从小麦7DL染色体上的长穗偃麦草衍生而来的抗叶锈病基因Lr19,用随机扩增多态性DNA(RAPD)和微卫星标记进行了标记。以抗叶锈病的Thatcher近等基因系(Tc + Lr19)与感叶锈病品系Agra Local杂交产生的340株F₂群体为作图群体,该群体对显性单基因叶锈病抗性进行了分离。分子标记在由140株幼苗组成的F₂衍生F₃纯合群体中进行定位。鉴定出16个RAPD标记与外源基因Lr19连锁,其中8个处于相引相连锁。12个RAPD标记与Lr19位点共分离。位于7D染色体长臂上的9个微卫星标记也被定位为与基因Lr19连锁,其中7个与Lr19位点共分离,从而在该位点两侧10.2 +/- 0.062 cM范围内产生了一个携带25个与基因Lr19连锁的分子标记的饱和区域。位于Lr19位点两侧的两个RAPD标记S265(512)和S253(737)分别转化为序列特征性扩增区域标记SCS265(512)和SCS253(736)。标记SCS265(512)与Lr19处于相引相连锁,标记SCS253(736)处于相斥相连锁,二者一起使用时可模拟一个共显性标记,能够区分杂合抗性幼苗和纯合抗性幼苗。这些分子标记在大多为Thatcher背景的近等基因系中进行了验证,这些近等基因系对44个不同的Lr基因(包括本地和外源来源)进行了等基因处理。对常见叶锈病感病品种多态性的验证也证实了这些与基因Lr19紧密连锁的标记在标记辅助选择中的实用性。