Direct development of plantlets from immature panicles of rice in vitro

Cultured immature panicles of rice formed plantlets from spikelets without callus or embryoid formation on MS and HE media containing 2 mg/l each of NAA and kinetin. Developmental stage, ploidy of explant and plant growth regulators in the medium are the major factors affecting the frequency of spikelet budding in young panicle culture. It is suggested that spikelet budding occurs by the reversion of floral primordia to vegetative stage or by the formation of adventitious buds from epidermal cells.     

Brassinosteroids function in spikelet differentiation and degeneration in rice

Brassinosteroids(BRs) play crucial roles in many aspects of plant development. However, their function in spikelet differentiation and degeneration in rice(Oryza sativa L.) remains unclear. Here, we investigated the roles of these phytohormones in spikelet development in fieldgrown rice subjected to five different nitrogen(N)fertilization treatments during panicle differentiation. BR levels and expression of genes involved in BR biosynthesis and signal transduction were measured in spikelets. Pollen fertility and the number of differentiated spikelets were closely associated with 24-epicastasterone(24-epiCS) and28-homobrassinolide(28-homoBL) levels in spikelets.Enhanced BR biosynthesis and signal transduction, in response to N treatment, enhanced spikelet differentiation, reduced spikelet degeneration, and increased grain yield. Increases in proton-pumping ATPase activity, ATPconcentration, energy charge, and antioxidant system(AOS) levels were consistent with 24-epiCS and28-homoBL concentrations. Exogenous application of24-epiCS or 28-homoBL on young panicles induced a marked increase in endogenous 24-epiCS or 28-homoBL levels, energy charge, AOS levels, spikelet differentiation, and panicle weight. The opposite effects were observed following treatment with a BR biosynthesis inhibitor. Our findings indicate that, in rice, BRs mediate the effects of N fertilization on spikelet development and play a role in promoting spikelet development through increasing AOS levels and energy charge during panicle development.     

Spikelet Sterility and Flowering Response of Rice to Water Stress at Anthesis

Water deficits at the anthesis stage of rice (Oryza sativa L.)induce a high percentage of spikelet sterility and reduce grainyield. This study attempted to elucidate the direct effectsof water stress on panicle exsertion, spikelet opening, andspikelet desiccation leading to spikelet sterility. A well-wateredtreatment and two water stress levels were imposed in pot-grownplants of IRAT 13 (upland cultivar) and IR20 (lowland cultivar)at the time of flowering under greenhouse conditions A cultivar difference was observed in the flowering responseto water stress with a high sensitivity in IR20. The time courseof panicle exsertion showed an inhibitory effect due to thelow panicle water status. Low panicle water potentials significantlyreduced the number of opened spikelets. Spikelet opening wascompletely inhibited at panicle water potentials below –1·8MPa and –2·3 MPa in IR20 and IRAT 13, respectively.However, the peak spikelet opening time in a day was not influencedby the stress treatment. Spikelets in stressed panicles wereobserved to remain open for a longer period than in the well-wateredpanicles. The role of turgor in spikelet opening is also discussedin the study. At low panicle water potentials, severe desiccationof spikelets and anthers was noted. The deleterious effectsof water deficits on spikelet opening and spikelet water losscontributed to reduced spikelet fertility Oryza sativa L., rice, spikelet sterility, flowering, water stress, panicle water potentials, turgor potentials, desiccation     

Polyamines and ethylene in rice young panicles in response to soil drought during panicle differentiation

This study tested the hypothesis that polyamines (PA) and ethylene (ETH) mediate the effects of soil drought on spikelet development in rice (Oryza sativa L.). Two rice cultivars, Yong You-2640 and Yang Dao-6, with vastly different panicle sizes were grown in pots under three soil moisture treatments: well-watered (WW), moderate soil drought (MD) and severe soil drought (SD), from the onset of panicle initiation to the pollen completion stage. MD treatment significantly increased spikelet differentiation, spikelet number per panicle, fully filled grain percentage and grain yield, decreasing the percentage of degenerated spikelets, sterile spikelets and partially filled grains compared to WW treatment. In contrast, SD treatment showed opposite effects. MD also increased the contents of free spermidine (Spd), free spermine (Spm) and the ratios of free putrescine, free-Spd and free-Spm to 1-aminocylopropane-1-carboxylic acid (ACC), decreasing the ETH evolution rate and ACC content in young panicles. In contrast, SD treatment showed opposite effects. Furthermore, free-Spd and free-Spm contents increased significantly, while ETH and ACC levels, and the percentage of degenerated and sterile spikelets decreased significantly under application of Spd or an inhibitor of ETH synthesis. The results were reversed when ACC or an inhibitor of Spd and Spm synthesis was applied. These findings suggest antagonistic interactions between free-PA (Spd and Spm) and ETH in response to soil drought, mediating spikelet development in rice.     

Over-expression of the rice LRK1 gene improves quantitative yield components

In rice ( Oryza sativa L.), the number of panicles, spikelets per panicle and grain weight are important components of grain yield. These characteristics are controlled by quantitative trait loci (QTLs) and are derived from variation inherent in crops. As a result of the complex genetic basis of these traits, only a few genes involved in their control have been cloned and characterized. We have previously map-cloned a gene cluster including eight leucine-rich repeat receptor-like kinase ( LRK ) genes in Dongxiang wild rice ( Oryza rufipogon Griff.), which increased the grain yield by 16%. In the present study, we characterized the LRK1 gene, which was contained in the donor parent (Dongxiang wild rice) genome and absent from the recurrent parent genome (Guichao2, Oryza sativa L. ssp. indica ). Our data showed that rice LRK1 is a plasma membrane protein expressed constitutively in leaves, young panicles, roots and culms. The over-expression of rice LRK1 results in increased panicles, spikelets per panicle, weight per grain and enhanced cellular proliferation, leading to a 27.09% increase in total grain yield per plant. The increased number of panicles and spikelets per panicle are associated with increased branch number. Our data suggest that rice LRK1 regulates rice branch number by enhancing cellular proliferation. The functional characterization of rice LRK1 facilitates an understanding of the mechanisms involved in cereal crop yield, and may have utility in improving grain yield in cereal crops.     

The LAX1 and FRIZZY PANICLE 2 genes determine the inflorescence architecture of rice by controlling rachis-branch and spikelet development

We have analyzed two mutants that exhibit altered panicle architecture in rice (Oryza sativa L.). In lax1-2, which is a new and stronger allele of the previously reported lax mutant, initiation and/or maintenance of rachis-branches, lateral spikelets, and terminal spikelets was severely prevented. In situ hybridization analysis using OSH1, a rice knotted1 (kn1) ortholog, confirmed the absence of lateral meristems in lax1-2 panicles. These defects indicate that the LAX1 gene is required for the initiation/maintenance of axillary meristems in the rice panicle. In addition to its role in forming lateral meristems, the wild-type LAX1 gene acts as a floral meristem identity gene which specifies the terminal spikelet meristem. A comparison of the defects in lax1-1 and lax1-2 plants suggested that the sensitivities to reduced LAX1 activity were not uniform among different types of meristems. In the fzp2 mutant panicle, the basic branching pattern of the panicle was indistinguishable from that of the wild type; however, specification of both terminal and lateral spikelet meristems was blocked, and sequential rounds of branching occurred at the point where the spikelet meristems are initiated in the wild-type panicle. This resulted in the generation of a panicle composed of excessive ramification of rachis-branches. The lax1-1 fzp2 double mutants exhibited a novel, basically additive, phenotype, which suggests that LAX1 and FZP2 function in genetically independent pathways.     

GENETIC ARCHITECTURE FOR THE ADAPTIVE ORIGIN OF ANNUAL WILD RICE, ORYZA NIVARA

The wild progenitors of cultivated rice, Oryza nivara and Oryza rufipogon , provide an experimental system for characterizing the genetic basis of adaptation. The evolution of annual O. nivara from a perennial ancestor resembling its sister species, O. rufipogon , was associated with an ecological shift from persistently wet to seasonally dry habitats. Here we report a quantitative trait locus (QTL) analysis of phenotypic differentiation in life history, mating system, and flowering time between O. nivara and O. rufipogon . The exponential distribution of effect sizes of QTL fits the prediction of a recently proposed population genetic model of adaptation. More than 80% of QTL alleles of O. nivara acted in the same direction of phenotypic evolution, suggesting that they were fixed under directional selection. The loss of photoperiod sensitivity, which might be essential to the survival of the ancestral populations of O. nivara in the new environment, was controlled by QTL of relatively large effect. Mating system evolution from cross- to self-fertilization through the modification of panicle and floral morphology was controlled by QTL of small-to-moderate effect. The lack of segregation of the recessive annual habit in the F₂ mapping populations suggested that the evolution of annual from perennial life form had a complex genetic basis. The study captured the genetic architecture for the adaptive origin of O. nivara and provides a foundation for rigorous experimental tests of population genetic theories of adaptation.     

Panicle Differentiation and Spikelet Number Related to Size of Panicle in Pennisetum americanum

The size of the developing panicle of pearl millet (Pennisetumamericanum (L.) Leeke) was studied during panicle differentiation(from panicle initiation to the completion of spikelet production)in plants grown in pots or in the field and supplied with varyinglevels of nitrogen. The duration of panicle differentiationrequired a constant thermal time (day degrees) under all nitrogensupplies. However, the rate of growth of the developing panicleduring this phase was retarded by low nitrogen supply. Duringpanicle differentiation, it appeared that the developing paniclehad to reach a critical size before developmental events suchas the initiation of spikelet primordia commenced; timing ofdevelopmental events was related to the size of the developingpanicle. The number of spikelets produced depended on the rate of growthof the differentiating panicle and the duration of the phaseof spikelet initiation (from appearance of the first spikeletprimordium to completion of spikelet differentiation). Low nitrogensupply reduced the number of spikelets produced, by retardingthe rate of growth of the differentiating panicle; this delayedthe time to initiation of spikelets and thereby reduced theduration of spikelet initiation. All spikelets (irrespectiveof nitrogen supply on the mainstem and on tillers) occupiedthe same area of panicle surface at the completion of differentiationof the panicle and at anthesis. Key words: Millet, Panicle differentiation, Spikelet number     

Genetic analysis of rice domestication syndrome with the wild annual species, Oryza nivara

With a small and sequenced genome, rice provides an excellent system for studying the genetics of cereal domestication. We conducted a quantitative trait locus (QTL) analysis of key domestication traits using an F2 population derived from a cross between the cultivated rice, Oryza sativa, and the annual wild species, O. nivara. We found that the QTL of large phenotypic effects were targeted by domestication selection for effective harvest and planting, including a reduction in seed shattering and seed dormancy and the synchronization of seed maturation. Selection for higher yield was probably responsible for the fixation of mutations at a cluster of QTL on chromosome 7 and a few other chromosomal locations that could have substantially improved plant architecture and panicle structure, resulting in fewer erect tillers and longer and more highly branched panicles in cultivated rice. In comparison with the wild perennial species, O. rufipogon, rice domestication from O. nivara would have involved QTL with a greater degree of chromosomal co-localization and required little genetic change associated with life history or mating system transitions. The genetic analyses of domestication traits with both wild relatives will open opportunities for the improvement of rice cultivars utilizing natural germplasm.     

Phenotypic plasticity in rice: responses to fertilization and inoculation with arbuscular mycorrhizal fungi

Aims Changes in the phenotype of crops (phenotypic plasticity) are known to play an important role in determining responses to nutrient availability, with the direction and magnitude of plasticity of individual traits being crucial for grain yields. Our study analysed the direction, magnitude and hierarchy of plastic responses of yield-related traits (i.e. biomass allocation and yield components) of rice (Oryza sativa L.) to nutrient availability. We estimated the effect of inoculation with arbuscular mycorrhizal fungi (AMF) on these characteristics of phenotypic plasticity.Methods A field experiment was carried out in northeast China, providing rice with six NPK fertilizer levels with or without inoculation with Glomus mosseae. At maturity, we quantified biomass allocation traits (shoot:root ratio and panicle:shoot ratio) and yield component traits (panicle number per hill, spikelet number per panicle, percentage of filled spikelets and seed weight). We also assessed the direction of change in each trait and the magnitude of trait plasticity.Important findings In non-inoculated plants, we found that biomass allocation and seed-number traits (i.e. panicle number per hill, spikelet number per panicle and percentage of filled spikelets) responded to fertilization in the same direction, increasing with rising fertilization. Panicle formation was the most plastic trait, while seed mass was the least plastic trait. AMF inoculation nullified the relationship between most biomass allocation and seed-number traits (except for that between panicle:shoot ratio and the percentage of filled spikelets) but increased the magnitude of plasticity in biomass allocation traits without altering the hierarchy of traits' plasticity. These results underscore the importance of plasticity of yield-related traits per se, and the impact of AMF on plasticity, for maintaining rice yields under low fertilization regimes.     

Comparative Proteomic Analysis Reveals Nitrogen Fertilizer Increases Spikelet Number per Panicle in Rice by Repressing Protein Degradation and 14-3-3 Proteins

The spikelet number per panicle is established in the early stages of panicle development. Nitrogen fertilizer application before panicle initiation is known to increase spikelet number, which is one of the most important traits in rice productivity determination. However, the basic proteomic mechanism remains poorly understood. The present study shows that nitrogen fertilizer significantly increased spikelet number and grain yield in rice. Proteomic variations were further analyzed in young panicles at the secondary panicle branch initiation and spikelet meristem initiation under nitrogen fertilizer treatment. Proteomic analysis identified 63 proteins with significant differential accumulation in young panicles under nitrogen fertilizer treatment. Proteolysis represents the largest functional category, which suggests that protein degradation is an important pathway in the response to nitrogen fertilizer. Importantly, nitrogen fertilizer significantly reduced 14-3-3 proteins, which interact with key enzymes associated with carbon and nitrogen metabolism, and the rice FT homologue Hd3a. Real-time PCR revealed that Hd3a signaling is also repressed by nitrogen fertilizer in leaves. This study contributes to a better understanding of the regulation of nitrogen fertilizers in the flowering pathway leading to panicle development. The identification of novel genes provides new insight into the profound impacts of nitrogen fertilizer on panicle development in rice.     

Involvement of 1-Methylcyclopropene in Plant Growth,Ethylene Production,and Synthase Activity of Inferior Spikelets in Hybrid Rice Differing in Panicle Architectures

Chlorophyll content (SPAD values) of leaves, 1-aminocyclopropane-1-carboxylic acid synthase (ACS) activities and ethylene production of inferior spikelets, aboveground biomass, grain yield, and yield components were studied to compare the effects of 1-methylcyclopropene (1-MCP) on plant growth, and inferior spikelet development in hybrid rice (Oryza sativa L.) differing in panicle architectures. There were four hybrid rice cultivars in this trial, including two lax-panicled cultivars, Liangyoupeijiu (LYPJ) and Guodao 6 (GD6), and two compact-panicled cultivars, Yongyou 9 (YY9) and Yongyou 12 (YY12). Results showed that chlorophyll content of leaves in 1-MCP treatment was higher than control (CK) after application of 1-MCP. ACS activities and ethylene production in 1-MCP treatment for LYPJ, GD6, and YY9 were significantly decreased compared to their corresponding CK. Aboveground biomass and grain yield of cultivars were increased with the increase of chlorophyll content of leaves, but decreased with the increase of ACS activities and ethylene production of inferior spikelets. On average, grain yield in 1-MCP treatment for LYPJ was increased by 7.3 % compared to CK, GD6 was increased by 2.6 %, YY9 was increased by 3.2 %, and YY12 was increased by 0.8 %, respectively. Application of 1-MCP showed marked effects on increasing spikelet fertility and harvest index of cultivars, but exhibited little effects on improving sink capacity and 1,000-grain weight. Our results indicate that 1-MCP could play a positive role in regulating the growth and development of hybrid rice with lax panicles or with compact panicles.     

Abscisic Acid and Ethylene Interact in Rice Spikelets in Response to Water Stress During Meiosis

This study investigated whether and how the interaction between abscisic acid (ABA) and ethylene is involved in the regulation of rice (Oryza sativa L.) spikelet sterility when subjected to water stress during meiosis. Two rice cultivars, HA-3 (drought-resistant) and WY-7 (drought-susceptible), were used and subjected to well-watered and water-stressed (WS) treatments during meiosis (15–2 days before heading). Leaf water potentials of both cultivars markedly decreased during the day as a result of the WS treatment, but panicle water potentials remained constant. The percentage of sterile spikelets in WS plants was increased by 49.7% for WJ-7 but only 12.7% for HA-3. ABA, ethylene, and 1-aminocyclopropane-1-carboxylic acid were all enhanced in spikelets by the water stress, but ethylene was enhanced more than ABA in WY-7 when compared with that in HA-3. Spikelet sterility was significantly reduced when ABA or amino-ethoxyvinylglycine, an inhibitor of ethylene synthesis, was applied to the panicles of WS plants at the early meiosis stage. Application of ethephon, an ethylene-releasing agent, or fluridone, an inhibitor of ABA synthesis, had the opposite effect, and sterility was increased. The results suggest that antagonistic interactions between ABA and ethylene may be involved in mediating the effect of water stress on spikelet fertility. A higher ratio of ABA to ethylene would be a physiologic trait of rice adaptation to water stress.     

近地层臭氧浓度升高对杂交稻颖花形成的影响

依托全球唯一的稻田开放式空气中臭氧浓度增高系统平台,以汕优63和两优培九为供试材料,设置大气背景臭氧浓度和高臭氧浓度（比大气背景臭氧浓度高50%）两个浓度水平,研究FACE条件下高O₃浓度对杂交稻颖花形成的影响.结果表明：高O₃浓度使汕优63和两优培九每穗颖花数分别减少28朵和34朵,下降幅度分别为15%和13%.从稻穗构成看,高O₃浓度胁迫下杂交稻每穗颖花数减少主要与每穗2次枝梗颖花数明显减少有关,对每穗1次枝梗颖花数的影响较小,因此高O₃浓度胁迫下水稻每穗1次枝梗颖花数占全穗的比率增加,每穗2次枝梗颖花数占全穗的比率降低.从颖花形成看,高O₃浓度胁迫下杂交稻每穗颖花数下降主要是颖花（特别是2次颖花）的分化受到抑制所致,而颖花的退化数不增反降.上述结果表明,采取相应措施削弱高O₃浓度胁迫对颖花分化的抑制作用可能是近地层高O₃浓度条件下减少杂交稻产量损失的关键.     

甲基磺酸乙酯(EMS)对水稻离体幼穗影响的初步研究

用不同浓度的EMS处理水稻离体幼穗,测定培养初期的呼吸强度、细胞色素氧化酶活性和同工酶以及苹果酸脱氢酶同工酶,发现低浓度的EMS对细胞呼吸强度以及细胞色素氧化酶活性有刺激作用,而高浓度的EMS则表现为抑制效应。呼吸强度和酶活性与幼穗培养力之间具有平行关系。研究结果表明,EMS的原初生理效应是影响幼穗细胞的代谢过程,幼穗体细胞诱导频率的改变是幼穗培养初期供能系统受到影响的结果。     

Genetic variation in the chloroplast genome suggests multiple domestication of cultivated Asian rice (Oryza sativa L.).

Two hundred and seventy-five accessions of cultivated Asian rice and 44 accessions of AA genome Oryza species were classified into 8 chloroplast (cp) genome types (A-H) based on insertion-deletion events at 3 regions (8K, 57K, and 76K) of the cp genome. The ancestral cp genome type was determined according to the frequency of occurrence in Oryza species and the likely evolution of the variable 57K region of the cp genome. When 2 nucleotide substitutions (AA or TT) were taken into account, these 8 cp types were subdivided into 11 cp types. Most indica cultivars had 1 of 3 cp genome types that were also identified in the wild relatives of rice, O. nivara and O. rufipogon, suggesting that the 3 indica cp types had evolved from distinct gene pools of the O. rufipogon - O. nivara complex. The majority of japonica cultivars had 1 of 3 different cp genome types. One of these 3 was identified in O. rufipogon, suggesting that at least 1 japonica type is derived from O. rufipogon with the same cp genome type. These results provide evidence to support a polyphyletic origin of cultivated Asian rice from at least 4 principal lineages in the O. rufipogon - O. nivara complex.