Genomic in situ hybridization analysis of Brassica napus × Orychophragmus violaceus hybrids and production of B. napus aneuploids

To further utilize the valuable germplasm Orychophragmus violaceus for Brassica genetics and breeding, a B. napus × O. violaceus cross was repeated with embryo rescue. All F₁ plants except one B. napus haploid were mixoploids (2n = 17–39 in ovaries) with 2n = 31, 37, 38 and 39 as the maximal chromosome numbers in individuals, but the higher numbers mostly appeared in pollen mother cells (PMCs) with a preponderance of 2n = 30, 37 and 38. Only one chromosome and one chromosome segment of O. violaceus were detected at a low frequency in some ovary cells and PMCs with 2n = 37, 38 and 39 as determined by genomic in situ hybridization analysis. The fatty acid profiles of seeds from the majority of the F₁ and F₂ plants were similar to those of female B. napus cv. ‘Oro’, but some were obviously different in the percentages of oleic, linoleic and erucic acids, and some F₂ plants (2n = 38) with good seed set had high percentages of oleic (>70.0%) or linoleic (to 38.3%) acids and low erucic acid (<1%). Subsequently, many kinds of B. napus aneuploids (2n = 28, 30, 34, 36, 37, 39 and 42), without O. violaceus chromosomes, were derived from F₂ progeny and microspores of partial F₁ plants. Finally, the cytological mechanisms behind the variations in chromosome numbers were discussed together with the implications of these aneuploids for Brassica genome research and of the plants with altered fatty acid profiles for improving the oil quality of B. napus.     

Establishment and characterization of Brassica juncea–Orychophragmus violaceus additions, substitutions and introgressions

Intergeneric hybridizations between cultivated Brassica species and Orychophragmus violaceus have been shown to be an efficient approach to produce Brassica aneuploids. Herein B. juncea–O. violaceus additions, substitutions and introgressions were obtained among progenies of partial hybrids after B. juncea (2n = 36) was pollinated by O. violaceus (2n = 24) and they expressed some traits from O. violaceus or novel for two parents. The single O. violaceus chromosome which was added to or substituted into the B. juncea chromosome complement was distinguishable most easily in pollen mother cells at anaphase I (AI) from its darker staining and more condensed form. The one pair of the O. violaceus chromosome in the substitutions paired and segregated regularly with others from B. juncea, and the single one formed one bivalent with one B. juncea chromosome and showed normal segregation and was more darkly stained in some AI group. Stable introgressions with 2n = 36 gave amplified fragment-length polymorphism (AFLP) profiles mainly similar to those of the female B. juncea parent, but fragments specific for O. violaceus could be found in every plants together with those deleted in B. juncea and novel bands for two parents. The mechanisms behind these unusual results are discussed.     

Meiosis and pollen grain viability in Helianthus mollis,Helianthus salicifolius,Helianthus maximiliani and their F1 hybrids with cultivated sunflower

Three diploid perennial sunflower species are useful for variety improvement: Helianthus mollis, because of sessile leaves, H. salicifolius, because of a high oil concentration, and H. maximiliani, a potential source of resistance to Sclerotinia sclerotiorum. The crossability of these species to cultivated sunflower was examined.Hybrids were obtained from eight combinations, with 3–15 F₁ plants per combination. The F₁'s exhibited the dominant phenotype of the wild species. Pollen viability varied between 32.1 and 69.9%. Meiosis was irregular in the F₁ hybrids. At diakinesis, bivalents (62.7–97.9% of meiocytes), univalents (0–31.23%), and multivalents (3.84–7.68%) were detected. At anaphase I, chromosome bridges were detected in 6.77 to 11.44% of meiocytes. Fast chromosomes in metaphase I, and lagging chromosomes in anaphase I and telophase II were evidenced in a high percentage of meiocytes.     

Somaclonal variation for some morphological traits, herb yield, essential oil content and essential oil composition in an Indian cultivar of rose-scented geranium

Somaclonal variation for plant height, plant spread, leaf shape, leaf size, leaf form, herb yield, essential oil content and 10 important constituents of the essential oil was studied in an Indian cultivar, ‘Bourbon’ of rose‐scented geranium (Pelargonium graveolens). Significantly larger variance was observed among in vitro‐regenerated plants of the SC1 generation (first generation following an in vitro phase) than among parental plants raised from stem cuttings for herb yield, plant height, leaf size, essential oil content, and for the contents of cis‐rose oxide, trans‐rose oxide, isomenthone and 10‐epi‐γ‐eudesmol in the essential oil. Plants of the SC₁ generation showing values beyond the higher or lower limit (depending on the trait) of the range observed among parental plants for different traits were developed into clones by their multiplication through stem cuttings, and evaluated again in the SC₂ generation (second generation following the in vitro phase). Somaclones selected for plant spread, and for the contents of linalool and trans‐rose oxide in their essential oils did not breed true in the SC₂ generation. However, for the remaining traits studied, 13–100% of the selected clones retained their selected traits in the SC₂ generation, suggesting that the changes in these somaclones were due to genetic causes.     

Cytogenetics and morphology of the pentaploid hybrid between Brassica napus and Orychophragmus violaceus and its progeny

A pentaploid hybrid plant (2n= 50, AACCO) between Brassica napus (AACC) and Orychophragmus violaceus (OO) showed matroclinous morphology and some patroclinous characters. Cloned progenies were mixoploid, consisting of various cells with 38–53 chromosomes, half of the cells with 50 chromosomes. The 50 chromosomes were mainly paired as 25 bivalents and segregated as 25:25 or 22:28; many other segregations were observed in some cells. Progenies produced by selfing had 38–47 chromosomes. Plants with 38 chromosomes were cytologically stable; in all other plants the chromosome number of individual cells varied between the genotype-specific maximum number and 38, indicating loss of chromosomes during mitosis. The mixoploid plants with 44 chromosomes mainly produced two kinds of mixoploid progenies with 44 and 41 chromosomes, respectively. All plants with 2n= 38 had the same morphology as the parental B. napus and were normal in fertility. These chromosome pairings and segregations in the pentaploid and its progeny support the hypothesis that O. violaceus is a natural polyploid species with a basic chromosome number of 3.     

芥菜型油菜和黑芥与诸葛菜属间杂种的获得及其特性

属间远缘杂交是创造新种质、解决育种材料短缺的一种有效方法。为了使诸葛菜的优良基因转入芥菜型油菜和黑芥中, 采用子房培养技术, 进行芥菜型油菜和黑芥与诸葛菜的正反属间杂交。在6种不同的培养基中培养子房。结果表明, 不同的杂交组合、培养基和母本的属间杂交可交配性不同。通过子房培养, 获得了10株属间杂种, 其中9株在形态上近似母本, 或介于父母本之间, 尤嫩斯与诸葛菜杂交组合的1株杂种表现出完全的雄性不育, 并在分枝类型和叶型方面表现出明显的变异; 形态学和SSR分子生物学鉴定表明10株杂种都是真正的属间杂种。     

A comparison of traditional and haploid-derived breeding populations of oilseed rape (Brassica napus L.) for fatty acid composition of the seed oil

Summary Microspore embryogenesis technology allows plant breeders to efficiently generate homozygous micros-pore-derived breeding populations of oilseed rape (Brassica napus L.) without traditional generations of inbreeding. This study was conducted to compare the frequency distribution of microspore-derived population and single seed descent populations with respect to fatty acids of seed oil. Both microspore-derived populations and single seed descent populations were produced from each of three crosses made between selected parents containing contrasting amount of erucic, oleic, linoleic and linolenic acids. The fatty acid content of F₃ plants derived lines (F₅ seed) developed by single seed descent was compared to that of microspore-derived populations. The means, ranges and distribution pattern of seed fatty acid contents were similar in both populations for each fatty acid studied, although a few heterozygous lines were observed in the single seed descent populations. The results indicated that microspore-derived population form random, homozygous F₁ plant derived gametic arrays for all fatty acids evaluated. Selection for altered fatty acid composition in microspore-derived and single seed descent homozygous populations should be equally efficient, in the absence of linkage of traits investigated.     

Selection for Seed Yield in Lolium perenne L.*

Components of seed yield were estimated for each set of 100 plants derived from six varieties of Lolium perenne. Significant differences between the varieties were found for all components of seed yield, but the range was broader within each variety. An indirect selection of seed yield using the character seeds per spike produced a selection response similar to that obtained by direct selection on high seed yield per plant. A direct and indirect selection of characters estimated in small, drilled plots of topcross off-springs (F₁) was less successful than the selection based on the single plants. Spearman rank correlation coefficients between the single plants and their offsprings did not show significant agreement for seed yield. But altogether the results indicated that the increase of seed yield/area was about 6 % when 10 % of the best plants were selected.     

Breeding effects on sensitivity of barley grain weight and quality to events of high temperature during grain filling

Monogenic inheritance and linkage were established on the basis of F₁ observations and analysis of 13,498 plants in 42 crosses for leaf colour, 7046 plants in 27 crosses for plant pubescence, 1926 plants in 8 crosses for number of leaflets per leaf, and 3182 plants in 12 crosses for plant height under field conditions. Normal green colour of foliage was found to be dominant over light green, pubescent plant over glabrous, high number of leaflets per leaf over low number of leaflets, and tall plant over dwarf. Linkage was estimated from joint segregation analysis, taking two characters at a time in all possible combinations as significant χ² values were recorded for these genes. Gene symbols Gl, Pub, Ph, and Hl are proposed for these four traits, respectively. The genes are arranged in the order of Ph-Gl-Pub-Hl with the map distances of 21.1, 28.9, and 17.5 cM between them. This short sequence of four linked genes spanning over 37.2 cM has been called Linkage Group 2 of lentil.     

Increasing biodiversity of irrigated rice in Africa by interspecific crossing of Oryza glaberrima (Steud.) × O. sativa indica (L.)

The African rice Oryza glaberrima, traditionally cultivated since more than 3.500 years, is of poor agronomic performance but resistant/tolerant to various stresses and diseases. The introduction of these characters into O. sativa cultivars is difficult since crossing barriers cause spikelet sterility in F1. Backcrossing can restore fertility and recently facilitated the development of fertile O. glaberrima × O. sativa ssp. japonica hybrid progenies for rain fed systems. With the objective to gain access to African rice germplasm for improvement of irrigated rice, crosses were performed with eighteen O. glaberrima and twenty O. sativa ssp. indica accessions. In total about one hundred F1-hybrid grains were obtained. The F1 plants were all completely sterile and backcrossing (BC) to O. sativa was performed in order to restore spikelet fertility. Monitoring of Tog5681 × IR64 hybrid progenies under field conditions revealed a broad genetic diversity within the BC₁ and BC₂ populations. Some BC₁ and BC₂ progeny plants outperformed the O. sativa parent, indicating that the heterozygocity level and complementary gene action after two backcrosses are still sufficient to positively influence plant vigor. Spikelet fertility of progenies was highly variable, but almost complete fertility was already observed within the BC₁F₂ population. High spikelet fertility was preserved in one out of two analyzed BC₁F₃ families and inmost of the BC₂F₃ families. The ability to restore spikelet fertility within few generations and the potential of the genetic diversity present in interspecific progenies facilitates the development of plant types specifically designed for the African irrigated and lowland environment. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetics of Resistance in Vicia sativa L. to Orobanche crenata Forsk.

The genetics of resistance of common vetch (Vicia sativa L.) to broomrape (Orobanche crenata Forsk.) was studied for two years by using the P₁, P₂, F₁, BC¹, BC₂, F₂ F₃, and F₄ generations obtained from crosses between resistant and susceptible lines. Resistant lines were selected by screening a world collection m a naturally infested plot. Resistance was tested both under field and greenhouse conditions. The best index to measure resistance was the number of emerged broomrapes per host plant. The results fit the additive-dominance model. The main component of the variation was additivity; dominance and interaction effects seemed to depend on the environment. When dominance is expressed, a low number is dominant over a high number of broomrapes per host plant.     

Impact of low linolenic acid content on seed yield of winter oilseed rape (Brassica napus L.)

An essential quality improvement of rapeseed oil can be obtained by reduction of its linolenic acid (C18:3) content from about 10% to less than 3% of the total fatty acids. Genotypes low in C18:3 have been developed by mutagenesis. The initial summer rapeseed mutant had been low yielding and highly susceptible to various diseases. It has been debated whether the low C18:3 character can be successfully combined with high seed yield for physiological reasons. Therefore, the low linolenic character of mutant M48 was transferred into high-yielding genotypes by repeated backcrossing to well-adapted low erucic acid, low glucosinolate (00-) winter rapeseed cultivars. Lines with low C18:3 content were selected from BC3 and BC4 generations and examined in 1990–95. Positive selection response for seed yield was shown to continue over the years. Presently, the best lines are yielding as well as the control cultivars being equivalent also in oil and glucosinolate contents. In order to test the effect of a low C18:3 content on seed yield, plants with low and with high C18:3 content, respectively, were selected from 16 segregating BC5-F₂ populations and bulked to form 32 F₃ populations. These ‘isogenic’ bulk populations were tested for field performance at four locations in 1995. The results show that C18:3 content of the seed oil is not associated with seed yield, oil content, beginning of flowering, plant height and disease resistance. Means of relative seed yield for the high and the low linolenic F₃ bulk populations were not significantly different with 88.0% and 86.9% of the control cultivars, respectively. There was a significant interaction between genotypes with high or low C18:3 content and location. This shows that under specific environmental conditions a low C18:3 content may be either favourable or unfavourable. The results indicate that the low C18:3 character of the original mutants per se does not cause a decrease in seed yield, oil content or general field performance.     

Anther culture response of wild Oryza species

Thirteen different wild species of the genus Oryza were investigated for their response to anther culture and plant regeneration. Callus induction from microspores of anthers was found to be strongly dependent on the species. Although large numbers of anthers from wild Oryza species, including O. barthii, O. latifolia, O. australiensis, O. minuta, O. nivara, O. paraguagensis and O. eichingeri, were cultured, no calli could be obtained. However, calli were produced from anthers of O. punctata, O. perennis, O. alta, O. ridleyi and O. rufipogon, although the frequency of callus induction was different. Similar species-dependence was observed in plant regeneration from microspore-derived calli. In total, 62 plants were derived from anther culture, including 47 albino and 15 green plants (of which 26.7% were haploids) from O. perennis; for the first time, six albino plants were obtained from O. ridleyi. Phytohormone combinations in the callus induction medium were found to influence callus induction and different wild Oryza species exhibited their own preferred phytohormone combinations for anther culture. In general, NK medium containing suitable concentrations of auxin and cytokinin may be successfully applied for anther culture of selected wild Oryza species.     

Development of a large population of activation-tagged mutants in an elite indica rice variety

In this study, we have generated more than 12,000 activation-tagged mutants in a high-yielding indica rice variety, 'BPT 5204', employing maize Ac/Ds system. Different transgenic plants obtained were analysed based on expression patterns of green fluorescence protein (GFP), red fluorescence protein (RFP), herbicide (Basta) tolerance and molecular analyses. T₁ seeds of pSQ5 and pSQ5-bar transgenics, when germinated separately on hygromycin (50 mg/L) and phosphinothricin (5 mg/L) containing medium, revealed a segregation of 3 tolerant : 1 susceptible plants. The germinal transposition frequency of Ds element in different T₂ progeny of rice plants was found to be about 18.0%. Different stable tagged mutants exhibited marked increases in plant height, number of tillers, leaf size, panicle size, seed size and number of grains per plant. The overall results indicate that the genes associated with these traits are upregulated by the enhancer element in activation-tagged mutants. As such, the various tagged mutant lines appear promising and serve as a valuable genetic resource for identification of key genes determining different agronomic traits of rice.     

Evaluation of somaclonal variation for genetic improvement of patchouli (<Emphasis Type="Italic">Pogostemon patchouli</Emphasis>), an exclusively vegetatively propagated aromatic plant

Patchouli (Pogostemon patchouli) is an important, exclusively vegetatively propagated aromatic plant, whose essential oil is widely used in perfumery and cosmetic products. Forty SC1 generation (first generation following in vitro phase) somaclones selected randomly from about 400 somaclones developed from the variety Johore, were multiplied through stem cuttings and evaluated in SC₂ and SC₃ generations to study the extent of somaclonal variation generated for plant height, herb yield, essential oil content, essential oil yield, and seven constituents of the essential oil. Significant or highly significant somaclonal variation was observed for plant height, herb yield, essential oil content, essential oil yield, and contents of patchouli alcohol, α-guaiene, α,δ-patchoulene, and α-bulnesene in the essential oil. The number of somaclones significantly superior to the parental variety for plant height, herb yield, essential oil content, and patchouli alcohol content in the essential oil ranged from 8–16 and the maximum superiority over the parental variety for these traits ranged from 21–79%. Broad-sense heritability estimates of plant height, herb yield, and essential oil content were 0.60–0.70 while those of essential oil yield and patchouli alcohol content were 0.44 and 0.47, respectively. Heritability estimates of other studied essential oil constituents were generally low (0.12–0.38). A high positive correlation was observed between essential oil yield and herb yield suggesting that selection for herb yield would be effective in improving essential oil yield. Patchouli alcohol content in the essential oil was negatively correlated with all the studied traits. Somaclonal variation, heritabilities of traits, and inter-trait correlations are reported for the first time in patchouli.     

Growth and Anti-Oxidative Systems Changes in Elymus dahuricus is Affected by Neotyphodium Endophyte Under Contrasting Water Availability

Although the relationship between fungal endophyte and agronomic grass (Lolium perenne and Festuca arundinacea) in drought tolerance are well documented, the mechanisms responsible for wild grass are not well understood. In this study, we determined the biomass production and growth parameters endophyte infected (E+) and endophyte uninfected (E−) Elymus dahuricus plants under high water (HW) and low water (LW) treatment for 8 weeks in a controlled‐environment condition experiment. We also determined the changes in the activities of the anti‐oxidative enzymes superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX) and content of H₂O₂, as well as levels of proline and chlorophyll a + b were examined in the leaves of E+ and E− plants. Under low water treatment, E+ plants produced more biomass and had higher values in plant height and tiller numbers, but no influence by the fungus were observed in high water treatment. Anti‐oxidative enzyme (include SOD, POD, CAT and APX) activities and contents of proline and chlorophyll a + b increased and H₂O₂ concentration declined in the E+ plants compared with E− plants under low water treatment. Therefore, endophyte infection was a benefit to growth and anti‐oxidative affection E. dahuricus under low water treatment.     

Genetic variability and interrelationship of traits in the industrial oil crop Vernonia galamensis

Vernonia galamensis is a wild plant from the family Asteraceae which is endemic to East Africa and has the potential to become a new oil crop for industrial uses. Its seed oil is rich in vernolic acid, a fatty acid of high interest for oleochemical applications. However, a breeding program for Vernonia galamensis cultivars with high seed and oil yields requires knowledge about the genetic variability of traits that influence seed and oil production. This study was undertaken to examine phenotypic and genotypic variability, broad-sense heritability, genetic advance under selection and interrelationships of agronomic and seed quality traits. A total of 122 Vernonia accessions, 115 collected from different regions of Ethiopia and seven introduced, were grown at two locations in Ethiopia (Alemaya and Babile), in 2001/2002 and were analyzed for 20 traits including phenology, yield, yield components, and seed quality with special emphasis on fatty acid composition. The collections exhibited significant variation for all traits except for days to emergence. Genotypes and locations interacted significantly (P 0.01) for all traits. Broad-sense heritability estimates ranged from 11% (for days to emergence) up to 79% (for days to maturity). Expected genetic advance was between 1.3% (for days to emergence) and 44.8% (for seed oil yield). Genetic correlation analysis revealed that seed yield per plant is highly and positively correlated with seed weight and head number; highly significant and negative correlations (r = –0.59, –0.82, –0.85, and –0.89) were found between vernolic acid and palmitic, stearic, oleic, and linoleic acid, respectively. Highly significant positive correlations (r = 0.55, 0.44, and 0.36) were observed between vernolic acid and oil content, meal protein content and seed oil yield, respectively. Path-coefficient analysis indicated seed weight and secondary head number to be the most important components of seed yield per plant. Vernolic acid, oleic acid and linoleic acid had positive direct effects and stearic acid had a negative direct effect on oil content. The direct positive effect of oleic acid on oil content was, however, compensated by the negative indirect effects of stearic and vernolic acid resulting in a negative correlation (r = –0.60) between oleic acid and oil content. These observations will support the selection of accessions with high seed and oil yield, high meal protein contents, and high vernolic acid content.     

Androgenetic response of heterozygous triticale populations using a greenhouse hydroponic system

Production of haploids, followed by chromosome doubling to produce doubled haploids (DH) represents the most rapid means of achieving complete inbreeding. In order to improve the androgenetic responses and maximize the production of green regenerants from selected plants of twenty-five triticale (× Triticosecale, Wittmack) populations (BC₁F₁, TC₁F₁, and F₂) we used a uniform and optimal growth environment for anther donor plants within a greenhouse hydroponic system. Non-orthogonal analysis of deviance showed highly significant differences (p < 0.001) among populations for both induction and regeneration. The overall induction response of the populations was very high with a mean of 50.4 embryoids per 100 anthers plated. Among all tested-populations, M86-6068/TW179//EP80 (TC₁F₁) was the most responsive for both induction and regeneration which could be associated to its Triticum timopheevii cytoplasm. On the other hand, although populations 80465/II83-194 both as BC₁F₁ and F₂ had a high level of induction response, only a few green plants were regenerated. These populations probably possess a partial Secale montanum genome, which could be contributing to the low regeneration ability. In conclusion, the need for optimization of donor plant growth conditions to effectively assess the androgenetic ability of individual populations/lines would be emphasized. This revised version was published online in August 2006 with corrections to the Cover Date.     

Apical Development and Growth of Barley under Different CO2 and Nitrogen Regimes

Increases in atmospheric carbon dioxide (CO₂) concentration have stimulated interest in the response of agricultural crops to elevated levels of CO₂. Several studies have addressed the response of C₃ cereals to CO₂, but the interactive effect of nutrient supply and CO₂ on apical development and spikelet set and survival has not been investigated thoroughly. Hence, an experiment was conducted in the greenhouse to evaluate the effect of high (700 μmol CO₂mol^?1 air) and low (400 μmol mol^?1) levels of atmospheric CO₂ on apical development, spikelet set and abortion, and pre- and post-anthesis growth in spring barley (Hordeum vulgare L.) grown under high N (0.3 g N pot^?1 before sowing ?1–0.11 g N pot^?1 week^?1) and low N (0.3 g N pot^?1) regimes. The plants were grown in 5 L pots. Development of spike was hastened due to CO₂ enrichment, and the C+ plants pollinated few days earlier than the C— plants. Carbon dioxide enrichment had no effect on date of ripening. Development of spike slowed following application of extra N, and plants pollinated 10 days later and matured 2 weeks later when compared with plants under low N. Carbon dioxide enrichment did not affect the number of spikelets at anthesis. Excess N decreased spikelet abortion and the increased maximum number of spikelets under both [CO₂]. Barley plants did not tiller when grown in low [CO₂] and low N. Increased endogenous IAA concentration in those plants, recorded three days before tillers appeared in other treatments, may have contributed to this. Carbon dioxide enrichment increased the C concentration of plants, but decreased the N concentration under high N regime. Both the C and N concentration of plants were increased under high N regime. Carbon dioxide enrichment increased the total dry matter of mature plants by 9 % under high N regime and by 21 % under low N regime. Under high [CO₂] increased kernel number on tiller spikes, and increased kernel weight both on main stem and on tiller spikes resulted in a 23 % increase in kernel yield under low N regime and 76 % increase in kernel yield under high N regime. The rate of N application influenced growth and yield components to a greater extent than CO₂ enrichment. At maturity, plant dry matter, kernel weight, the number of kernels per spike, and the number of spikes per plant were higher under high N regime than under low N regime. Long days (16 h), low light intensity (280 μmol m^?2s^?1), and at constant temperature of 20 °C high [CO₂] increased kernel weight and the number of kernels on tiller spikes under high and low N application rate, but did not increase the number of kernels on main stem spike, or the number of tillers or tiller spikes per plant.     

Modification of plant height via RNAi suppression of OsGA20ox2 gene in rice

GA 20-oxidase (GA20ox) is a regulatory enzyme for the syntheses of biologically active GAs in plants. The loss-of-function mutations in OsGA20ox2 of rice (Oryza sativa L.) generate the well-known Green Revolution gene sd-1, which cause the semi-dwarfism phenotype. In our present investigation, semi-dwarf plants were generated from a taller rice variety QX1 by RNAi suppression on the expression of OsGA20ox2. The 531bp-fragment of OsGA20ox2 was amplified by PCR from genomic DNA of QX1 and used to construct the hairpin RNAi vector pCQK2. The wild type QX1 was transformed with pCQK2 by Agrobacterium-mediated transformation and some independent transgenic RNAi lines exhibited semi-dwarfism. RT-PCR and Northern blot analyses showed that the expression of OsGA20ox2 was specifically suppressed in the RNAi semi-dwarf lines. Endogenous GA assays revealed that the contents of the GA20ox2-catalyzed products GA₁₉, GA₂₀ and the down-stream biologically active GA₁ were drastically reduced in the RNAi semi-dwarf lines. We further showed that the RNAi semi-dwarf lines could be restored to normal plant height by applying exogenous GA₃. The results indicated that the semi-dwarfism of the RNAi semi-dwarf lines was associated with the decreased expression of OsGA20ox2 gene and the reduced content of endogenous biologically active GA₁. Analyses of panicle length, seeds per panicle and 1000-grain weight suggested that the RNAi semi-dwarf lines showed stable grain yield compared with the wild type plants. It is demonstrated that the RNAi approach could be useful for plant breeding purposes in the future. Qing Yang, Chun-Lian Wang have contributed equally to this work.