The amylose contents of rice determine eating quality which is one of the major traits in rice breeding program. To identify the low-amylose gene of the japonica rice cultivar Baegjinju, genetic analysis was conducted using 200 F2 population derived from a cross between the japonica cultivars, Saeilmi and Baegjinju. Individual F2 plants were classified as wild type (translucent grain) and mutant type (dull grain) based on the grain appearance of brown rice. Two hundred F2 plants were segregated into 155 wild type plants and 45 mutant type plants, which fit the 3:1 ratio (x2 = 0.667, df = 1, p = 0.414) and this result indicated the low-amylose gene of Baegjinju is a single recessive gene which controls the amylose contents. Linkage analysis was conducted to localize the low-amylose gene of Baegjinju and fine mapped within an 800-kb interval between 17.5 to 18.8Mb on short arm of chromosome 10. Co-segregated SSR marker, RM25648 was developed and it could be useful for marker-assisted selection and determination of the genetic resource related with amylose contents in rice breeding.
Bakanae (foolish seedling) disease caused by Gibberella fujikuroi creates serious problems in the foremost rice growing countries. This study was conducted to identify new resistance genetic sources to Bakanae disease. Bioassay showed that 11 varieties including Gwangmyeongbyeo, Hawn, Wonseadaesoo, Erguailai etc. were resistant to bakanae disease among 254 rice germplasm. Mismatch ratio between phenotype on bakanae disease bioassay and allele type of RM9, a SSR marker closely linked the bakanae disease resistant QTL, qBK1, were 38.3%. These results suggest that RM9 might be used for selecting qBK1, but it cannot be used for wide range of rice germplasm. Resistant germplasm in this study might be have resistant genes different from qBK1. The eleven varieties resistant to selected in this study will be used to identify new resistant alleles or genes to improve bakanae disease resistance in rice.
Drought caused by global climate change is one of serious problems for rice cultivation. However, it was little reported the impact of drought on rice cultivation in Korea. In here, to assess impact of drought on rice varieties in Korean climate condition, growth characteristics and yield components of rice were compared on irrigated and partially irrigated rice paddy field. First, we have chosen 11 rice varieties including ‘Saeilmi’ and ‘Shindongjin’ which are widely cultivated in Korea. For partially irrigated rice paddy treatment, we have withheld irrigation from 25 days after transplanting and water supply was totally dependent on rainfall for rice cultivation. When we examined early plant height and tiller number of these varieties on partially irrigated rice paddy were reduced 1.6% to 18.4% and 10.4% to 33.1%, respectively, and these reduction rate were highly correlated with yield loss in our experimental conditions. Among rice yield components, panicle number was decreased 10.5% to 30.1% according to rice varieties and reduced panicle number was highly correlated with yield loss. Grain number per panicle, grain filling rate and 1,000 seeds weight did not have correlation with yield loss of rice varieties. These result means that growth stage, especially the tillering stage, is seriously affected by drought on rice cultivation in Korea. And we suggest that ‘Saeilmi’, ‘Ilmi’ and ‘Ilpum’ are good for rice cultivation on drought prone rice field in Korea.
In here, we screened drought tolerant varieties with modified leaf water loss rate assay and visual drought tolerant phenotype in the greenhouse conditions with more than 800 varieties. Among these varieties, Samgang, Gumei4 and Apo showed the lowest of leaf water loss rate and strong drought tolerant phenotype. To identify drought QTLs with Samgang variety, we developed the doubled-haploid (DH) population consist of 101 lines derived from a cross the drought tolerant cultivar Samgang and the drought sensitive cultivar Nagdong. To score the drought phenotype degrees of this population, we withheld water for 6 weeks and treated the watering for 7 days. After watering, visual phenotype was observed 1 to 9 degree according to the standard evaluation system for rice, IRRI. Drought sensitive parent Nagdong was almost died and was scored as 9 degree, while tolerant parent Samgang showed slightly leaf tip drying phenotype and was scored as 3 degree in our experimental conditions. Three main QTLs were detected on chromosome 2, 6, and 11 with this visual phenotype. We also measured relative water contend of these population under drought stress conditions, and got one main QTL on chromosome 11. The QTL loci on chromosome 11 with flanking markers RM26755-RM287 has a function for visual phenotype and relative water content under drought conditions.
Rice is a staple food crop for more than half of the world population. Severe losses of rice production was caused by various environmental conditions such as cold, heat and flooding annually. Rice is a highly sensitive to low temperature below 15-20 ℃ because of originating from tropical or subtropical climates. Especially, seedling of rice is easily damaged to low temperature and result in seedling yellowing, growth retardation, reduced tillering and yield losses at last. We used a recombinant inbreeding lines (RIL) population of 384 individuals derived from a cross between Hanareum 2, a highly cold sensitive variety and Unkwang, a cold tolerant variety for molecular mapping of QTLs related to cold tolerance. Seedling discoloration of each lines and parents caused by cold response were investigated in field condition after transplanting. And leaf samples of RIL population were collected for evaluation of chlorophyll content using 80% acetone extraction. The seedling of each lines and parents was subjected to low temperate by 5~13 ℃ during 14 days. The cold recovery score (CRS) of RILs was recorded after 4 days recovery period according to standard evaluation system (SES, IRRI). Total of eight QTLs were detected on chromosome 1, 7, 8, 10, 11 and 12 using cold tolerance traits, chlorophyll content, seedling discoloration and cold recovery score in 384 RILs. The qCRS12, which detected on chromosome 12 between two flanking markers id12002113, id12002563 (1.1 Mbp) showed 25 LOD score with 26% of phenotypic variation of cold recovery score in RILs population. The positive allele contributing to cold tolerance came from the cold tolerant parent Unkwang. The result may provide useful information for a marker-assisted breeding program to improve cold tolerant in rice.
Green rice leafhopper(GRH), Nephotettix cincticeps Uhier, is one of the major insect pests of rice (Oryza sativa L.) in the temperate growing region of East Asia. GRH sucks sap from both xylem and phoem of susceptible rice varieties, and increased GRH populations cause sooty mold disease on the ears of rice after heading stage. In addition to direct plant destruction, GRH also causes damage to rice plants by transmitting rice dwarf viruses causing rice dwarf viruses disease which could decrease the yield of rice. Development of GRH resistant rice varieties for reducing yield loss is an important objective in current breeding programs. In this study, we developed three SSR markers(RM18166, RM516, RM18171) and one Indel marker(Indel15040) which could select Grh1-resistant varieties using population derived from cross lines between Grh1-resistant variety ‘Singwang’ which contains Grh1 gene and susceptible variety ‘Ilpum’. PCR products of RM18166 which was one of the developed markers were easily detected in agarose gel. These markers will be useful for development of the Grh1-resistant varieties through marker-assisted selection(MAS) without bio-examination in rice breeding
Bakanae disease is one of the most serious and oldest problems of rice production, which was first described in 1828 in Japan (Ito and Kimura 1931). This disease may infect rice plants from the pre-emergence stage to the mature stage, with severe infection of rice seeds resulting poor germination or withering (Iqbal et al. 2011). Under favorable environmental conditions, infected plants have the capacity to produce numerous conidia that subsequently infect proximate healthy plants, resulting in major yield loss (Ou 1985). One hundred sixty nine NILs, YR28297 (BC6F4) generated by five backcrosses of Shingwang with the genetic background of susceptible japonica variety, Ilpum were used for QTL analysis. Rice bakanae disease pathogen, CF283, was mainly used in this study and inoculation and evaluation of bakanae disease was performed with the method of the large-scale screening method developed by Kim et al. (2014). A major QTL for resistance against bakanae disease on chromosome 1 was identified using SSR marker, RM9, which explaining 65 % of the total phenotype variation. The major QTL designated as qBK1 and mapped to a 4.4 Mbp region between RM24 (19.30 Mb) and RM11295 (23.72 Mb). The results of this study are expected to provide useful information toward developing resistant rice lines to this detrimental fungal disease.
Bakanae disease incidence threat is an increasing trend in the top rice growing countries. Despite it is essential to identify the resistant genes and underlying mechanisms of bakanae disease to develop resistant varieties, there are very limited genetic studies on bakanae disease in rice. The indica rice variety Shingwang was selected as resistant donor to bakanae disease. One hundred sixty nine NILs, YR28297 (BC6F4) generated by five backcrosses of Shingwang with the genetic background of susceptible japonica variety, Ilpum were used for QTL analysis. Rice bakanae disease pathogen, CF283, was mainly used in this study and inoculation and evaluation of bakanae disease was performed with the method of the large-scale screening method developed by Kim et al. (2014). The proportion of healthy plants of Shingwang and Ilpum after inoculation was confirmed using bakanae disease pathogen, CF283. While inoculated Ilpum showed thin and yellowish-green phenotype which is typical symptom of Bakanae disease, Shingwang showed similar healthy phenotype with control plants. A major QTL for resistance against bakanae disease on chromosome 1 was identified using SSR marker, RM9, which explaining 65 % of the total phenotype variation. The major QTL designated as qBK1 and mapped to a 4.4 Mbp region between RM24 (19.30 Mb) and RM11295 (23.72 Mb). The information of qBK1 could be useful for improving rice bakanae disease resistance in marker-assisted breeding.
The use of functional markers, it is expected to make direct identification about genetic diversity at DNA level and overcome the problem of recombination /linkage. These markers can be used to identify interesting alleles in a breeding program and indirectly select for the trait, saving money, time and labor. Bacterial blight of rice caused by Xanthomonas oryzaepv. Oryzae is a destructive disease in rice production worldwide. No bactericide is effective to control the bacterial blight disease yet. Xa3, which is a gene conferring resistance to BB of the rice plant has been previously characterized by map-based cloning. We have cloned and sequenced the Xa3/xa3 gene in Korean cultivar, Hwayoung, Ilmi and Goun with gene specific primers. Our work detected polymorphisms and PCR-based allele specific SNP markers were developed. Susceptible or resistant individuals from an F2 population developed from across between Milyang244 and Ilmi, Korean germplasms and near isogenic lines carrying BB resistance genes were screened with allele specific markers. We found that the genotype completely matched their phenotype to BB using ASP-primers. These markers could be effective to marker-assisted selection for the Xa3 gene in rice breeding programs.
The green rice leafhopper (GRH), Nephotettix cincticeps Uhler, is one of the most serious insect pests affecting cultivated rice (Oryza sativa L.) in temperate regions of East Asia. To understand the genetic basis of the GRH resistance, a F2 population derived from across between a highly resistant variety,Cheongnam and a susceptible variety, Junambyeo was analyzed by genetic analysis and association mapping. GRH resistance was evaluated using the F2 populations. The results showed that a single dominant gene in Cheongnam. DNA from 22 F2 individuals being either resistant or susceptible were pooled to produce bulk resistant and bulk susceptible DNA samples. Parents and bulks were screened with 192 SSR markers and twolinked SSRmarker, RM6082 and RM20145 were identified.Subsequent mapping in the original mapping population showed that thelocusis flanked by the SSR markers, RM20130 and RM20152 on chromosome 6. To physically map this locus, the-linked markers were landed on the artificial chromosome clones of the reference cv., Nipponbare, released by the International Rice Genome Sequencing Project. The DNA markers found to be closely linked to Grh3 would be useful for marker-assisted selection for the improvement of resistance to GRH in rice.
Amylose content of rice endosperm is one of the determinants of rice eating quality. This study was conducted to elucidate the mode of inheritance of dull gene in Milyang262, tentatively designated as du7(t), and to identify the molecular marker for du7(t) to be employed in marker-assisted breeding and gene pyramiding. Genetic analysis was carried out on F2 population derived from a cross between Junam and Milyang262. The low amylose content of Milyang262 was indicated to be under single recessive control. Allelism tests were as well conducted by crossing Milyang262 with Baegjinju and Baegokchal, which harbor du1 and wx gene, respectively. du7(t) was demonstrated to be inherited independently to du1 and wx. F2 population of Baegokchal/Milyang262 was used for molecular mapping. Linkage analysis was conducted on a population consisted of 120 individuals by several SSR markers. Initial mapping indicated that du7(t) is located on the end of long arm of chromosome 6 between SSR marker RM20590 and RM3509. To fine map the gene, a bigger population and several additional markers were employed. du7(t) was further mapped to a 1.74 Mb region between two SSR markers (RM6926 and RM412). Furthermore, we indentified three SSR markers that co-segregated with du(t) i.e. RM6811, RM3765, and RM176.
A phosphate starvation-induced acid phosphatase cDNA was cloned from the rice, Oryza sativa. The cDNA encoding O. sativa acid phosphatase (OsACP1) has 1100 bp with an open reading frame of 274 amino acid residues. The deduced amino acid sequence of OsACP1 cDNA showed 53% identity to tomato acid phosphatase and 46-50% identity to several other plant phosphatases. OsACP1 expression was up-regulated in the rice plant and in cell culture in the absence of phosphate (Pi). The induced expression of OsACP1 was a specific response to Pi starvation, and was not affected by the deprivation of other nutrients. OsACP1 expression was responsive to the level of Pi supply, with transcripts of OsACP1 being abundant in Pi-deprived root. The OsACP1 cDNA was expressed as a 30 kDa polypeptide in baculovirus-infected insect Sf9 cells. In addition, the OsACP1 gene was introduced into Arabidopsis via Agrobacterium mediated transformation. Functional expression of the OsACP1 gene in the transgenic Arabidopsis lines was confirmed by Northern blot and Western blot analyses, as well as phosphatase activity assays. These results suggest that the OsACP1 gene can be used to develop new transgenic dicotyledonous plants able to adapt to Pi-deficient conditions.