We have developed a polymerase chain reaction (PCR)-based assay that could effectively reduce the time period required to screen and select for Gall Midgeresistant rice lines under field conditions. The primers for the assay were designed on the basis of sequence information of two phenotype specific random amplified polymorphic DNA fragments which were found to be tightly linked to Gall Midge biotype-1 resistance gene (Gm2). The two RAPD fragments, F81700 in the susceptible parent 'ARC6650' and F10600 in the resistant parent 'Phalguna', were identified after screening 5450 loci using 520 random primers on genomic DNAs of 'ARC6650' and 'Phalguna'. These primers, when used in a multiplexed PCR, amplified specifically a 1.7-kb and 0.6-kb fragment in the susceptible and resistant parents, respectively. When this assay was performed on genomic DNAs of 44 recombinant inbred lines derived from 'ARC6650' x 'Phalguna' and 5 lines derived from other crosses where one of the parents was 'Phalguna', 'ARC6650' or their derivatives, the primers amplified a 1.7-kb fragment in all of the susceptible lines or a 0.6-kb fragment in all of the resistant ones. These markers can be of potential use in the marker-aided selection of Gall Midge biotype-1 resistant phenotypes. As screening for resistance can now be conducted independent of the availability of insects, the breeding of resistant varieties can be hastened.
Gm2 is dominant gene conferring resistance to biotype 1 of gall midge (Orseolia oryzae Wood-Mason), the major dipteran pest of rice. The gene was mapped by restriction fragment length polymorphism (RFLP) analysis of a set of 40 recombinant inbred lines derived from a cross between the resistant variety 'Phalguna' and the susceptible landrace 'ARC 6650'. The gene is located on chromosome 4 at a position 1.3 cM from marker RG329 and 3.4 cM from RG476. Since the low (28%) polymorphism of this indica x indica cross hindered full coverage of the genome with RFLP markers, the mapping was checked by random amplified polymorphic DNA (RAPD)/bulked segregant analysis. Through the use of 160 RAPD primers, the number of polymorphic markers was increased from 43 to 231. Two RAPD primers amplified loci that co-segregated with resistance/susceptibility. RFLP mapping of these loci showed that they are located 0.7 cM and 2.0 cM from RG476, confirming the location of Gm2 in this region of chromosome 4. Use of these DNA markers will accelerate breeding for gall midge resistance by permitting selection of the Gm2 gene independently of the availability of the insect.
Forty-seven recombinant inbred (RI) lines derived from a cross between two indica rices, cv 'Phalguna' and the Assam land race ARC 6650, were subjected to restriction fragment length polymorphism (RFLP) analysis using cloned probes defining 150 single-copy loci uniformly dispersed on the 12 chromosomes of rice. Of the probes tested, 47 detected polymorphism between the parents. Heterozygosity was calculated for each line and for each of the polymorphic loci. Average heterozygosity per line was 9.6% but was excessive (>20%) in the 5 lines that seemed to have undergone outcrossing immediately prior to harvest. Average heterozygosity detected by each probe across the 47 RI lines was 9.7%. The majority of probes revealed the low level of heterozygosity (<8%) expected for F5-F6 lines in a species showing about 5% outbreeding. On the other hand, 7 probes exhibited heterozygosity in excess of 15%, while with a eighth probe (RG2 from chromosome 11) heterozygosity varied according to the restriction enzyme employed, ranging from 2% with SaII to 72% with EcoRV. The presence of 34 recombination sites in a segment of the genome as short as 24 kb indicates a strong selection for recombination between two neighbouring loci, one required as homozygous for the 'Phalguna' allele, and the other heterozygous. Since selection was principally for yield advantage over that of the high-yielding parent, 'Phalguna', one or both of these loci may be important for heterosis in this cross. The results also indicate that heterozygosity as measured by RFLP can depend on the particular restriction endonuclease employed.
Genetics of tolerance to iron chlorosis was investigated in eight crosses involving parents distinctly different in their level of tolerance. The segregating populations with parents and F~s were screened under actual stress conditions in the field. Also, selected crosses were studied for Fe 3+ uptake capacity. Tolerance/moderate tolerance to Fe chlorosis was dominant over susceptibility and it was controlled by two sets of nonallelic genes with complementary interaction. Gene lc~ has been found to be basic and in complementation with Ic 3 it confers tolerance. Likewise, Ic 2 with Ic 4 confers tolerance. The basic genes lc~ and Ic 2 are nonallelic and, in the absence of their respective complementary genes Ic 3 and Ic 4, ineffective, which results in susceptibility. Of tolerant cultivars, ARC 10372 and Cauvery have been tentatively assigned the genotype of Ic~, ic2, Ic3, ic4, and moderately tolerant lET 7613, Prasanna and Akashi ic~, lc 2 Ic 3 Ic 4. The susceptible ARC 5723 has been assigned ic~, ic 2 Ic3, IC4, and lET 9829, ic~, ic 2 lc3, ic 4. lET 7614 is susceptible, due to the presence of inhibitory genes I-lc~, I-Ic 2 together with icl, ic2, Ic3, lc 4. Further, the gene Pc for purple coleoptile shows linkage with one of the complementary genes with a crossover value of 15.26%, while the gene(s) for seedling height "Is with Ic~ with a crossover value of 1.7%. It is possible that the gene(s) for iron chlorosis tolerance might belong to the second linkage group, where genes for purple leaf were located.
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