Agro-morphological characterization of ovary culture-derived plants of rice (Oryza sativa L.)

Plants derived from unpollinated ovary culture of ten rice genotypes showed significant variability in agro-morphological characteristics. The ovary-derived plant (H1) populations were completely haploid, doubled haploid or haploid-doubled haploid mixture. Haploids had very drastic reduction in plant height, panicle length, grain length, breadth and number and spikelet fertility (0.0%–2.1%). Doubled haploids from the hybrid of UPRI 95–121 × UPRI 95–165 were normal with fertility ranging between 69.6% and 97.7%. A genetic segregation in ratio of 1:1 was observed for five pigmentation characters in the H1 population derived from hybrid UPRI 95–122 × UPRI 95–165. Plant height showed the largest coefficient of variability (28.5%) followed by the number of spikelets per panicle (24.2%), number of grains per panicle (22.0%), percent seed set (9.2%) and panicle length (9.0%). The range of variation in the H1 population from fully fertile hybrid PMS 2A (CMS) × IR 31802 (restorer) was similar to its corresponding F2 population for plant height, spikelet fertility and number of grains/panicle. A single clone of plants from the cultivar BG 1321 exhibited complete male sterility but normal female fertility when pollinated with other varieties. Ovary-derived plants from the CMS lines PMS 2A and IR 58025A showed stable male sterility and those from thermosensitive genetic male sterile line UPRI 95–140 showed thermosensitive genetic male sterility. These lines have potential in the hybrid breeding program and are being currently exploited. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chromosomal regions controlling anther culturability in rice (Oryza sativa L.)

Diallel analysis has revealed that anther culturability in rice (Oryza sativa L.) is a quantitative trait controlled by the nuclear genome. Mapping of anther culturability is important to increase the efficiency for green plant regeneration from microspores. In the previous study, we detected distorted segregation of RFLP markers in rice populations derived from the anther culture of an F₁ hybrid between a japonica cultivar ‘Nipponbare’ and an indica cultivar ‘Milyang 23’. To clarify the association between chromosomal regions showing distorted segregation and anther culturability, the anther culturability of doubled haploid lines derived from the same cross combination was examined, and the association between alleles of the RFLP markers, which exhibiting the most distorted segregation on chromosomes 1, 3, 7, 10 and 11, and the anther culturability was evaluated. One region on chromosome 1 was found to control callus formation from microspores, and one region on chromosome 10 appeared to control the ratio of green to albino regenerated plants. In both regions, the Nipponbare allele had positive effects. Three regions on chromosomes 3, 7 and 11, however, showed no significant effect on anther culturability. This revised version was published online in August 2006 with corrections to the Cover Date.     

Complex constitutive nature of Japanese upland rice, Oryza sativa L.

Two rice ecotypes, the so-called lowland and upland populations, which carry different isozyme genotypes mostly at a single locus, are cultivated in Japan. The aim of this study was to examine the origin and the mechanism for keeping these genetic differences. The upland population is cultivated in upland fields and carries a different allele for a particular isozyme gene, Pgd-1, which has never been found in the lowland population. RFLP markers showed a weak trend for genetic differentiation between the two ecotypes. On the other hand, morphological, and physiological traits showed marked differences between the two ecotypes. Furthermore, based on the genotypic difference, two Japonica subgroups are defined in the upland population. Subgroup I is the minor group and carries key lowland characters, including the genotype for PGD. Subgroup II carries different traits and the genotype for PGD of the alternative subgroup. As an allelic difference for Pgd-1 is known to occur between the two ecospecies, Tropical (Tr) and Temperate (Tm) Japonicas, upland cultivars can be classified by diagnostic characters which distinguish a variety into Tr or Tm type. The upland population consists of three types of cultivars, Tr-, Tm- and intermediate-type. In contrast, the lowland population consists of a uniform Tm type Japonicas. As Japanese upland cultivars still have an isozyme allele specific to the Tr type, the upland population has a rather complex constitution which is presumably now being introgressed by lowland genetic material, but still represents a major difference at some genetic levels. Upland rice carries several stress-resistant genes which would be useful for lowland rice breeding. The genetic difference would be efficient for tagging upland specific traits by upland specific genetic markers. This revised version was published online in July 2006 with corrections to the Cover Date.     

High-lysine mutants of rice, Oryza sativa L.

In order to improve the nutritive value of rice protein, it is necessary to increase the level of lysine and threonine. Ten high-lysine mutants were selected from N-methyl-N-nitrosourea-induced (MNU-induced) mutant lines, using high-performance liquid chromatography for amino acid analysis. The lysine content of these mutants ranged from 5.10% to 6.38% of total amino acids, while that of the original varieties was about 4%. All high-lysine mutants also had a greatly increased histidine content and a floury endosperm. An increase in lysine content of the albumin/globulin fraction was characteristic of the MNU-induced high-lysine mutants. The segregation ratio of normal to floury endosperm in the F2 seeds from a cross between ‘Kinmaze’ and a high-lysine mutant fitted the expected 3:1 ratio. The high level of lysine and the floury endosperm may be controlled by the same gene.     

Callus Formation and Plant Regeneration from Protoplasts Derived from Suspension Culture of Rice (Oryza sativa L. cv. 'Taipei 177')

Protoplasts were isolated from young inflorescence-derived suspension cultures of a japonica rice cultivar ‘Taipei 177’. The isolated protoplasts which were cultured either in liquid, agar on Sea plaque agarose underwent sustained division. Maximum plating efficiency of 1.06% occurred in a medium containing macroelements of KM, microelements and vitamins of B5, 0.5 % Sea plaque agarose, 1.0 mg/l of 2,4-D, and glucose as an osmotic stabilizer. Green and albino plants were regenerated from the protocalli in MS semisolid medium containing 4 mg/l BAP, 0.5 mg/l NAA and 500 mg/l casein hydrolysate (MS_18–2).     

Breeding of widely compatible restorer lines in rice (Oryza sativa L.) through anther culture

Following anther culture of various F₁ hybrids of indica restorer (R) lines/wide compatible varieties (WCVs) and japonica R lines/WCVs, the homozygous diploid plants (2n) generated were test-crossed with indica WA type cytoplasmic-genetic male sterile (CMS) line Zhanshan 97A (WA), indica testers Nanjing 11 and Nante, japonica BT CMS line Hanfeng A (BT), and japonica testers Balilla and Akihikari to identify widely compatible restorer lines. The results of this study showed that among the diploid pollen plants generated from F₁ hybrids of indica R lines/WCVs, 36.7% and 64.7% possessed normal fertility restoration ability (rate of seed-setting > 80%) to Hanfeng A and Zhanshan 97A, respectively. 71.3% and 32.3% had normal fertility restoration ability to Hanfeng A and Zhanshan 97A, respectively, in diploid pollen plants derived from the japonica R lines/WCVs F₁'s anther culture. Several widely compatible R lines were selected from anther culture of F₁ hybrids of indica R lines/WCVs and japonica R lines/WCVs. These widely compatible R lines derived from diploid pollen plants showed good wide compatibility and restoration ability both to WA and BT type CMS lines. Strong standard heterosis of major agronomic traits and yield traits was observed in F₁ test-crosses of widely compatible R lines with Hanfeng A and Zhanshan 97A. The wide compatibility in widely compatible R lines H17, H158 and H281 was studied by a set of three-way crosses. Segregation of the fertile plants with seed-setting rate > 70% and semi-fertile plants with seed-setting rate > 69.9% agreed with a ratio of 1:1, indicating that wide compatibility in these widely compatible R lines is governed by a pair of major genes. This revised version was published online in July 2006 with corrections to the Cover Date.     

A Functional Growth Analysis Model for Grain Development in Oryza sativa L.

The pattern of grain growth in two rice cultivars was assessed in terms of total dry weight of grain from anthesis upto maturity. Rice grain development followed a sigmoidal trend. Richards' function was found to more closely fie the data on grain growth and thus could serve as a suitable mathematical model for rice grain growth analysis.     

Enhanced anther culture efficiency of indica rice (Oryza sativa L.) through modification of the culture media

The production of microspore-derived green plants from anther culture of indica rice is generally very low compared with japonica cultivars. A modified anther culture medium, consisting of a higher KNO₃ content (31 mm ) and casein hydrolysate (CH, 500 mg/1) but without ammonium salts, was tested in comparison with a medium consisting of the widely-used N6 medium nitrogen background, using four indica × indica F₁ hybrids as test materials. Green plant regeneration frequency was at least three-fold higher in the microspore-calli derived from the former medium than in those derived from the modified N6 medium. More than 700 microspore-derived plants were raised in the field. Another study was carried out using indica × japonica and indica × javanica F₁ hybrids. The results indicated that a medium with higher (3.5 mm ) ammonium sulphate may induce a higher frequency of anthers with microspore-calli but not necessarily lead to a larger number of green-plant regenerating calli. Subsequently, using the indica cv. ‘IR-43’ as the test material, use of a lower level (1.75 mm ) of (NH₄)₂SO₄, in addition to KNO₃ (31 mm ), was found to be better than CH (500 mg/l) for anther-response as well as green plant regenerability of the derived microspore-calli. Nitrate-nitrogen or ammonium-nitrogen alone elicited poor response. Twenty-five media involving combinations of KNO₃ (20–34 mm ) and (NH₄)₂SO₄ (1–3 mm ) were tested for their effects on anther response. Combinations involving KNO₃ (31–34 mM) and (NH₄)₂SO₄ (2.0–2.5 mm ) were found superior not only for achieving greater anther response but also, for subsequent green-plant regeneration. This contrasts with the 28 mm of KNO₃ and 3.5 mm of (NH₄)₂SO₄ in the widely-used N6 medium developed for japonica rice. Other than potassium nitrate and ammonium sulphate, and potassium phosphate to some extent, the levels of other inorganic salts tested did not make any significant difference to the process of anther response. Based on these results, modified media with three levels of ammonium sulphate were tested for anther culture efficiency of indica × japonica and indica × javanica derivatives (F₃s). Microspore-calli derived from a medium of a lower (1.75 mm ) level of (NH₄)₂SO₄ showed a higher regeneration potential overall than those derived from a higher (3.5 mm ) level. A revised medium has been suggested, on the basis of these results, for the realization of improved anther culture efficiency and, consequently, improved feasibility of using doubled haploids in genetic and breeding research with indica rice.     

粳稻和籼稻愈伤诱导和分化期间生理代谢差异初探

通过比较粳稻品种日本晴和籼稻品种9311成熟胚愈伤诱导和分化阶段,愈伤组织的基础生理代谢差异,探讨籼稻9311再生率低于粳稻日本晴的可能原因。研究表明,在愈伤诱导和分化的所有不同时期,在所有时期日本晴愈伤中的总氮和氨态氮含量均高于9311,在分化25天以前,硝态氮含量也高于9311。此外,在细胞代谢水平上,通过对可溶性蛋白含量的差异分析表明,在愈伤诱导一个月至分化15天时,日本晴的可溶性蛋白含量高于9311。     

Screening of Photoinsensitive Summer Rice (Oryza sativa L.) Genotypes for Ratoon Cropping

Of the 24 genotypes screened for ratooning ability and ratoon grain yield, only ten showed regeneration. RP 1664-4461 had the highest regeneration. Ratoon growth duration varied from 49–61 days. Ratoon grain yield was highest (1.7 t/ha) for RP 1664-4461 and the lowest (0.8 t/ha) for IET 7613. Productivity per day in the ratoon crop was comparable to that of the main crop. Based on grain yield and per day productivity of main and ratoon crops, cultures CR 222 MW 10, Rasi, RP 1664-4461, RP 1165-132-1 and CR 215-55-54-1 appeared promising for ratoon cropping.     

Inheritance of photoperiod sensitive genic male sterility and breeding of photoperiod sensitive genic male-sterile lines in rice (Oryza sativa L.) through anther culture

The fertility segregations of F₁, F₂, BCF₁ descended from crosses between PSGMR and japonica varieties, and F₁'s anther cultured homozygous diploid pollen plant populations (H₂) were studied to reveal the genetic mechanism of photoperiod sensitive genic male sterility in PSGMR under natural daylight length at Shanghai. Rate of bagged seed-setting was used as an indicator of fertility. Fifteen F₁ showed complete fertility similar to their parents. The ratio of completely sterile plants to fertile plants in fifteen F₂ and four BCF₁ was 1:15 and 1:3, respectively. The ratio of completely sterile to fertile diploid pollen plants in nine diploid populations (H₂) was 1:3. These results demonstrated that the photoperiod sensitive genic male sterility in PSGMR was governed by two pairs of independent major recessive genes. There were no significant fertility segregations in hybrids F₁ and selfed F₂ between Nongken 58S and its derivatives 7001S, 5088S, 5047S and M105-9S, indicating that the photoperiod sensitive genic male-sterile genes in Nongken 58S were allelic to those in its derivatives. Several photoperiod sensitive genic male-sterile diploid pollen lines were bred from anther cultured homozygous diploid populations (H₂) in about a three-year period. Most of these diploid lines showed significant fertility transformation and stable complete sterility from 5 August to 5 September, excellent agronomic traits and high resistance to blast and bacterial leaf blight. This revised version was published online in July 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.     

The use of microsatellite polymorphisms for the identification of Australian breeding lines of rice (Oryza sativa L.)

Ten microsatellite loci were analysed for 43 cultivars or breeding lines of rice. Polymorphism-Information-Content values ranged from 0.62 to 0.92. The microsatellite markers were found to be useful for cultivar identification and assessment of genetic relationships. Most of the cultivars could be uniquely identified by at least one microsatellite marker. Genetic heterogeneity was detected within rice samples by amplification of microsatellites from DNA extracted from multiple individual plants and also from bulked DNA preparations. This revised version was published online in July 2006 with corrections to the Cover Date.     

水稻营养生长发育阶段系统的划分

文章综述了水稻营养器官（根、茎、叶）生长阶段的系统划分。在以往形态学和解剖学阶段划分的基础上,整合了近年来分子遗传学的研究进展,突变体的发现使发育阶段的划分更加准确和完善。从分子水平搞清水稻的结构及发育阶段可以为水稻的高产和稳产提供理论基础。     

Ratoon Growth and Development in Rice (Oryza sativa L.) Under Various Temperature Regimes

Plants exhibited differential response for ratoon growth and development under different temperature regimes before and after main crop harvest. Low temperature (23°/20 °C) before cutting induced ratooning whereas similar temperature after cutting delayed and reduced ratoon tillering. Ratoon grain yield and its attributes were significantly affected under various temperature regimes. Ratoon grain yield under low temperature regime before cutting did not differ from that of the control. Ratoon growth duration was maximum under low temperature regime after cutting. High temperature (37°/27 °C) after main crop cutting resulted in high spikelet sterility in the ratoon crop.     

Genetic progress of upland rice (Oryza sativa L.) lines for disease resistance

The rice crop is affected by diseases throughout its cycle, impacting negatively on grain yield and quality. The control of the disease impact can be accomplished via crop breeding, using highly multiple resistant genotypes. This study aimed to evaluate the efficiency of multiple-character and specific selection of multiple resistance to major culture-associated diseases (neck blast, leaf scald and grain discoloration) in rice lines of the Upland Rice Genetic Breeding Program. The experiments were conducted in 35 sites during 12 agricultural years, where 124 lines were evaluated for the severity of fungal diseases, under natural field conditions. Multiple parameters were calculated based on the diseases´ scores: genetic, phenotypic and environmental variances, heritability, selection gain, renewal rate, and genetic and renewal progress. Genetic variance for the disease resistance was identified in the population, and the selection gain for multiple-character selection was of 3.16 year⁻¹ throughout the breeding process with a renewal rate of over 35%. The programme has showed efficiency in selecting multiple resistant genotypes to the mentioned diseases, highlighting genotypes with high potential for market release.     

Trisomic analysis of a strong photoperiod-sensitivity gene E1 in rice (Oryza sativa L.)

Recent genetic analyses on heading-time of rice indicated that almost all the well-adapted varieties in the temperate zone carry a strong photoperiod-sensitivity gene E1, a dominant allele of E1 locus. In order to identify the chromosome on which E1 is located, a trisomic analysis was made using two primary trisomic series originating from the japonica varieties, Nipponbare and Kinmaze, respectively. The Nipponbare and Kinmaze series were crossed with heading-time tester lines, EG0 and EG3, respectively, both of which did not carry the E1. The F2 populations for chromosome 1, 2, and 3 could not be analyzed due to lack of seed. All the other F2 populations showed distinct segregation into early-type and late-type plants caused by the E1 locus segregation, which suggested that the trisomic analysis for E1 locus could be efficiently made. Both disomic and trisomic groups in the F2 population from the cross of the trisomic line for chromosome 7 × EG0 showed a segregation ratio significantly different at the 1% level from a ratio of 1 [e1e1; early]: 3 [E1e1, E1E1; late]. This suggested that E1 was located on chromosome 7. Subsequently, the linkage analysis was made using three morphological marker genes on chromosome 7. It was recognized that E1 was linked to rfs (rolled fine strip gene) and slg (slender glume gene) with recombination values of 16.3 ± 5.88 and 9.1 ± 4.72%, respectively. From these results, it can be concluded that E1 is most likely to be located on chromosome 7. This revised version was published online in July 2006 with corrections to the Cover Date.