Anther culture of tropical japonica × indica hybrids of rice (Oryza sativa L.)

Summary Nine japonica × indica F1 hybrids of rice involving 6 indica and 3 japonica tropical varieties, were large scale anther cultured. The frequency of callusing anthers averaged 18.7%. The microspore-derived calli produced green plants with a mean frequency of 8.7%. Albino plants represented 61% of the shoot forming calli. Monitoring of the green and albino plant regenerating capabilities of calli arising between week 4 and week 8 of incubation of the anthers showed no increase of the albino/green ratio and a slow decrease of the shoot forming ability of the transferred calli after the sixth week of culture. Spontaneous doubled haploids (SDH) represented 46% of the regenerated green plants in 4 hybrids. However, a high frequency of partially sterile regenerants was noticed among 132 SDH plants generated from a hybrid.     

Haploidy breeding and mutagenesis for drought tolerance in wheat

Several intraspecific crosses between known drought tolerant wheat varieties and stable high yielding recombinants were made with the objective to develop improved cultivars for the moisture stressed rainfed areas of Pakistan. Five of these crosses were selected for further creation of useful mutations through the application of low doses of gamma rays and development of doubled haploids through anther culture. Anther culture response of the selected irradiated F₁ generations was studied on liquid and solid induction media. The highest number of calli among almost all crosses was produced on Potato-2induction medium. All the crosses varied greatly in response to callus induction and maximum calli (75%) were obtained from Lyl-73/vee’s’ cross. Similarly, genotypic differences were found for green vs. albino regenerants. The highest number of green plantlets (12.1%) was recorded for Lu-26/3062. From the developed doubled haploid population 25 DH-mutants were initially selected and nine lines were finally included in multi-locational field tests. Two DH-mutants (i.e. DHML-50 and DHML-9) have potential for better grain yield, earliness, disease resistance and moisture stress tolerance. This revised version was published online in August 2006 with corrections to the Cover Date.     

Anther and pollen grain culture of rice (Oryza sativa L.)

Summary Factors favouring callus proliferation and subsequent regeneration of plants from pollen grains of rice anthers (Oryza sativa L., cvx. Bahia, Girona, Balilla × Sollana and Sequial) were determined. Cultivar differences in response were found, such as a high rate of haploid plant regeneration.In addition, isolated pollen grain culture was used to induce tissue proliferation outside the anther walls. The frequency of callus formation from isolated pollen grains was very low. It was necessary to preculture the anthers before pollen grain separation, in order to accomplish a successful development later. Root differentiation was observed in some of the obtained callus.     

Production of salt tolerant rice breeding line via doubled haploid

Summary A haploid breeding program was initiated to develop doubled haploid salt tolerant rice breeding line via anther culture. Two sensitive breeding lines BR4608-R₁-R₂ and BR4909-R₁-R₂ were crossed with a salt tolerant line IR13146-13-3-3 to transfer its salt tolerant character to the doubled haploids.Anther from confirmed F₁s of the two crosses were cultured in defined medium for callus induction and eventual plant regeneration. Fifteen doubled haploid (DH) lines were obtained from two crosses. Test for salt tolerance were done in vitro. Five out of 15 lines were found tolerant at the level of 8–10 decisiemens/m (ds/m) while the rests were sensitive to that level of salinity.Field experiment was conducted to evaluate the doubled haploids under saline and non saline soil. Five salt tolerant lines produced comparable yield with the resistant control (BR 23) under saline condition, whereas these lines yielded even higher in non saline soil under irrigated condition when evaluated with other 10 sensitive DH linesAbbreviations LSD Least Significant Difference - NAA Napthalene Acetic Acid     

QTL mapping and associated marker selection for the efficacy of green plant regeneration in anther culture of rice

Anther culturability of rice is a quantitative trait controlled by nuclear‐encoded genes. The identification of quantitative trait loci (QTL) and associated marker selection for anther culturability is important for increasing the efficiency of green plant regeneration from microspores. QTL associated with the capacity for green plant regeneration in anther culture of rice were mapped on chromosomes 3 and 10 using 164 recombinant inbred (RI) lines from a cross between ‘Milyang 23’ and ‘Gihobyeo’. The quantitative trait locus located on chromosome 10 was detected repeatedly when three anther culture methods were applied and was tightly linked to the markers, RG323, RG241 and RZ400. Associations between these markers and the efficacy of green plant regeneration in 43 rice cultivars and two F₂ populations, ‘MG RI036’/‘Milyang 23’, and ‘MG RI036’;/‘IR 36’ were analysed. One of these markers, RZ400, was able to identify effectively genotypes with good (> 10.0%) and poor (< 3.0%) regenerability, based on the marker genotypes in the cultivars and two F₂ populations. This marker enables the screening of rice germplasm for anther culturability and introgression into elite lines in breeding programmes.     

Genetic analysis of male sterility in rice mutants with environmentally influenced levels of fertility

Summary Twenty-six male sterile plants grown in the field were recovered in the M7 generation from ethyl methane sulfonate-treated material of the rice cultivar M-201. Fertility increased five-fold when ratooned plants from the field were grown in a growth chamber with a 12 hour daylength. Crosses between mutant and normal fertile cultivars produced fertile F₁ plants. Female fertility was normal as judged by percent seed set from unbagged panicles of parental and recombinant lines. Transgressive segregation for fertility was observed for all crosses in the F₂ and F₃ generations. Five of 37 F₃ male sterile plants showed moderate levels of seed fertility under winter greenhouse conditions and reduced seed set when transplanted to summer field plots. Fertility data from reciprocal crosses suggested cytoplasmic factors had little or no effect on levels of male sterility in the mutant lines. Chi-squared analyses of F₂ and F₃ generation results indicated male sterility of the mutants is conditioned by two nuclear genes with epistatic effects.     

Induction of mutations for heat tolerance in potato by using in vitro culture and radiation

Heat tolerant mutants were obtained in two commercial potato cultivars, `Kufri Jyoti' and `Kufri Chandramukhi' through in vitro mutagenesis of in vitro propagated plantlets. Gamma-irradiated (20 and 40 Gy) shoots were micropropagated for three cycles (M₁V₃). A large number of the micropropagated shoots produced microtubers at 28 ^°C. Microtubers induced at high temperature had distorted shape but showed normal germination in field. Under stress conditions of high temperature, the frequency of chlorophyll variants increased in the gamma irradiation-derived material, however, nearly 40% of the plants had normal leaf tissue, whereas control plants showed completely damaged leaves. This revised version was published online in July 2006 with corrections to the Cover Date.     

Specific developmental stages of gametogenesis for radiosensitivity and mutagenesis in rice

Summary Developmental stages during gametogenesis of rice were histologically examined in the period from differentiation of reproductive organs to anthesis. Plants were exposed to acute X-rays of 20 Gy. Radiosensitivity and mutation frequency were investigated in relation to the developmental stages of reproductive organs. The most radiosensitive stage, as measured by reduction of the M1 pollen-and seed-fertilities, was the last premeiotic interphase. Mutations induced at different developmental stages were scored in M3 strains. Sterility mutants and short-culm mutants were most frequently observed. Grain shape, panicle morphology, heading-date and endosperm character mutants were induced at a relatively low frequency. The overall mutation frequency varied with the developmental stage at the time of irradiation. The highest overall mutation frequency was observed when radiation was applied 10 days before anthesis, the late tetrad stage of microspores. Radiation exposure of florets at the late tetrad stage was found to be a more efficient method of inducing a large number of mutations than radiations applied to seeds and fertilized egg cells.     

Inheritance of aroma in rice

Summary Inheritance of an aroma was worked out in crosses between Brimful from Nepal as an aromatic rice and leading Japanese varieties Koshihikari and Nipponbare as non-aromatic ones. The F₂ pattern of segregation for aroma to non-aroma was 3:13 indicating one dose gene for aroma and one dose inhibitor gene in two crosses. This ratio was confirmed by genetic behavior of F₃ populations.     

Inheritance of grain threshability in rice

Summary Inheritance of grain threshability was studied in a cross between two indica rice varieties. Hard threshability was found to be completely dominant over easy threshability. The segregation pattern indicated threshability to be monogenically controlled.     

Inheritance of salt tolerance in rice

Summary The genetic behavior of salt tolerance was studied in artificially salinized conditions at the International Rice Research Institute.Divergent selection, carried out at a salinity level where the ECe was 15.2 mmhos/cm at 25 C in F₃ lines from two crosses confirmed the effects of salt tolerance on F₄ progeny with realized heritability values of 0.39 and 0.62, respectively.In a cross between two tolerant cultivars there was clear over-dominance for tolerance, despite the high environmental fluctuation which resulted in a low genetic response as indicated by a low but significant repeatability of 0.20–0.25, and many progeny lines more tolerant than the parents were recovered. The superior tolerance of these progenies compared to the parents was confirmed subsequently at 3 different salt levels. In the same experiment a cross between tolerant and susceptible cultivars produced some progeny of comparable tolerance with tolerant sources.In a 6×6 diallel cross experiment with two tolerant, moderate, and susceptible varieties each, both general and specific combining ability were significant.The findings indicate the possibility of breeding rices more tolerant than existing tolerant cultivars through cumulative crosses of tolerant cultivars. Further improvement can be attained by crossing highly tolerant lines with donors of good agronomic traits and pest and disease resistance.     

Studies on the amylose content and gelatinisation temperature in certain local cultivars and induced grain shape mutants in rice

Summary Ninety local cultivars and 124 induced grain shape mutants were screened for their amylose content and gelatinisation temperature as indexed by the alkali disintegration patterns. The amylose content was high in most of the local cultivars and ranged from 19.2 to 32.0% and from 12.3 to 33.2% in the mutants. Wide variation was also observed for the alkali disintegration values and most of the varieties had low alkali values. A positive but low correlation between alkali values and amylose content and a negative correlation between alkali values and protein content was observed. The preliminary studies suggest that the amylose content at different milling levels increased with an increase in the level of milling. Some of the local cultivars and induced mutants were found to be superior with high amylose and high gelatinisation temperature and could be utilized in breeding programmes directed towards improving the cooking quality of high yielding varieties.     

Inheritance of scent in rice

Summary Inheritance of scent in rice and its association with pigmented characters were studied in T-141 × K-44-1. Four complementary genes appeared to control scent, one of them showing linkage with a complementary gene for glume colour and a supplementary gene for red pigment in apiculus. This group formed part of V (Prp)linkage group of Misro et al. (1966) in which case Pg and P were additions.Pl.480 Project on linkage maps in rice.     

Two loosely linked genes controlling the female specificity for cross-incompatibility in rice

Cross-incompatibility caused by endosperm abortion was found in advanced generations of backcrossing between the Asian wild (W593) and cultivated (T65wx) rice strains. The near isogenic line, T65WxS ₆ (W593), carrying a segment of chromosome 6 from W593 showed a low seed setting when pollinated with pollen grains of T65wx in spite of the fact that the reciprocal cross gave a high seed setting. The unidirectional or asymmetric crossing barrier was previously explained by an interaction between Cif and cim, both of which acted sporophytically, resulting in the cross-incompatibility reactions in the female (CIF) and male (CIM), respectively. In the genetic model, endosperm abortion is induced only when CIF gametes are fertilized with CIM gametes. This predicted that the double homozygote for Cif and cim might be self-incompatible since the plant expresses both CIF and CIM simultaneously. However, we failed to obtain such a self-incompatible plant by transferring Cif into a cim plant. The present results showed that CIF is controlled not only by Cif but also by an additional gene(s) loosely linked with it. We propose here that Cif ₁ (formerly named as Cif) and Cif ₂ determine CIF. In addition, Cif ₂ and Cim were not separated due to restriction of recombination, which might explain why it is difficult to obtain a self-incompatible rice plant expressing both CIF and CIM.     

水稻花器官数目突变体mf2的鉴定和基因定位

水稻的花器官发育影响着水稻的产量与品质。本研究通过¹²C重离子诱变航恢7号获得一个水稻花器官突变体multi-floret 2 (mf2), 其稃片、浆片、雄蕊、雌蕊增多, 多数小穗内具2~3朵类似小花。mf2内外稃不能很好勾合, 而且形状和维管束的数目都产生了一定程度的变化。电镜扫描幼穗发现花器官的变异在幼穗分化期的各花器官原基分化时就已形成。另外, 该突变体的抽穗期推迟, 株高降低, 穗数增多, 表明其营养生长也受到一定的影响。遗传分析表明mf2突变体表型受单隐性核基因控制。利用SSR、InDel分子标记将MF2定位于第1染色体的标记SSR39108和InD39210之间, 区间大小约为102 kb。     

Combination of stem elongation ability with submergence tolerance in rice

Summary A study was conducted with six F₂ populations to test the possibility of combining submergence tolerance and stem elongation ability into a single genotype of rice (Oryza sativa L.). Submergence tolerance and stem elongation ability could be combined in the same genotype if strongly submergence tolerant genes are present in submergence tolerant parents.     

Identification of QTLs controlling rice drought tolerance at seedling stage in hydroponic culture

‘Drought avoidance’ and ‘drought tolerance’ are two mechanisms by which plants adapt under water stress. These mechanisms are difficult to evaluate separately in field experiments. Using hydroponic culture, we studied the genetic control of drought tolerance in rice (Oryza sativa L.) without the effect of drought avoidance. A backcross inbred population of ‘Akihikari’ (lowland cultivar) × ‘IRAT109’ (upland cultivar) with 106 lines was cultured with (stressed condition) and without (non-stressed condition) polyethylene glycol (PEG) at seedling stage. The relative growth rate (RGR), specific water use (SWU), and water use efficiency (WUE) showed significant genotype × environment interactions with or without PEG, indicating that each line responded differently to water stress. A quantitative trait locus (QTL) analysis revealed that these interactions were QTL specific. A total of three QTLs on chromosomes 2, 4, and 7 were detected for RGR. The QTL on chromosome 7 had a constant effect across environments, while the QTL on chromosome 4 had an effect only under non-stressed condition and that on chromosome 2 only under stressed condition. The stress-specific QTL on chromosome 2 was not co-located with any QTLs for root system depth previously reported from the same mapping population. However, this QTL was co-located with a stress-specific QTL for SWU, suggesting that the control of transpiration was relevant to dry matter production under drought. We concluded that PEG-treated hydroponic culture is very effective for use in genetic analyses of drought tolerance at seedling stage.     

Genetic base of Japonica rice varieties released in Taiwan

Summary The pedigrees of 99 Japonica rice (Oryza sativa L.) varieties released between 1940 and 1987 were traced back to 65 ancestors, of which 44 were plant introductions from Japan and only 11 from Taiwan. Japanese introductions collectively contributed more than 85% of the parentage of the varieties. Although more ancestors were integrated into recent breeding programs, as few as 10 ancestors comprised 66% of the genetic background. Extensive use of superior genotypes from the same sources explained why the integration of new germplasm did not result in the diversification of the released varieties. The 10 most important ancestral contributors of the 99 varieties were from Japan except Oloan-chu from Taiwan. Two introductions, Sinriki and Kameji, had the highest mean relative genetic contributions of the genes with 21.3% and 16.7%, respectively. Shinriki was the most frequently used ancestor and occurred in 83 of the 99 pedigrees. These results clearly revealed the narrow genetic base in current Japonica rice varieties of Taiwan.     

A novel environment-induced genic male sterile (EGMS) mutant in indica rice

Summary Male-sterile mutants were isolated from M₂ and M₃ generations of indica rice variety 26 Zhaizao, dry seeds of which had been exposed to ⁶⁰Co- rays at a dose of 290 Gy. The mutants were planted in early season and ratooned in late season for two successive years for identification of fertility conversion in different growing seasons. One of the mutant lines was further observed in a growth chamber and in the field. Results showed that daily average temperature might be the major factor conditioning the male fertility conversion at a moderate daylength. The critical temperature for the male fertility conversion of the mutant grown under 12.5 h and 14.0 h daylength is about 23°C, below which the plant becomes completely male sterile. Its male fertility conversion character differs from other EGMS lines so far developed. The performance of the hybrids between the mutant and some other indica varieties demonstrated its good combining ability and its potential value in hybrid rice production. The obtained mutant line still sheds KI-stainable pollen grains under male sterilizing conditions. Nevertheless, pollen grains shed from the male sterile plants were much more vulnerable than from normal plants. At sucrose concentration below 1.5 M, the pollen grains from the mutant grown under male sterilizing conditions almost completely broke down, while above 1.5 M they became plasmolysed and shrunken. This is indicative of poor development of the membrane and walls of the pollen grains from the male sterile mutant, causing the pollen grains to be unfunctional. NBT test also clarified the abortion of the pollen grains from the mutant, which were formed in the male sterilizing environment.     

Genetic analysis of the response to day length in rice

Summary Inheritance of time-to-flowering, plant height, tiller number, spikelet number and flag leaf area was studied under day lengths of 10 and 14 hours in a diallel cross of six rice varieties. Early flowering was dominant to late in both environments but the varieties flowering early in one environment were late flowering in the other. Analysis of F₁ and F₂ data from the cross of Heenati-310 x IR-8 suggested a digenic control of early flowering in short days with complementary interaction. It is considered that while relatively few genes control time-to-flowering in rice a previous proposal that separate genes for time-to-flowering and photoperiod sensitivity exist is unnecessary on present evidence. Whereas culm length, tiller number and flag leaf area were increased by longer days, the number of spikelets per panicle was reduced. The length of the panicle was little affected by changing environment, and throughout certain characters and varieties, such as tiller number in I-geo-tze, were more stable than others. Varietal crosses of Heenati-310 x IR-8 and Tainan x MI-273(m) appeared to give high yield potential.