Comparative Study of CaMsrB2 Gene Containing Drought‐Tolerant Transgenic Rice (Oryza sativa L.) and Non‐Transgenic Counterpart

Concerns regarding the safety of transgenic foods have been raised because of possibility of undesirable effects development during genetic engineering. Analysis of phenotypic traits can increase the likelihoods of identifying those unintended effects in dietary composition of the GM crops. Objective of this study was to compare the transgenic lines with their non‐transgenic counterpart. Different vegetative and reproductive traits as well as antioxidant properties were considered to evaluate the transgenic (HV8 and HV23) lines containing CaMsrB2 gene and their non‐transgenic (Ilmi) parent line. Grain size and weight, seed germination, root length, root and shoot dry weight, length and width of flag leaf, plant height, and ligule, stamen and carpel length were not significantly different. Onset and completion of heading in each line occurred almost during the same period. The antioxidant properties in terms of DPPH (1,1‐diphenyl‐2‐picrylhydrazyl) radical scavenging activity and polyphenol content were not statistically different under same treatment condition. The results suggested that the transgenic rice lines containing CaMsrB2 gene were equivalent to their non‐transgenic counterpart without any visible unintended effects.     

Multiplex PCR genotyping for five bacterial blight resistance genes applied to marker‐assisted selection in rice (Oryza sativa)

Bacterial blight (BB) is the most economically damaging disease of rice in Asia and other parts of the world. In this study, a multiplex PCR genotyping method was developed to simultaneously identify genotypes of five BB resistance genes, Xa4, xa5, Xa7, xa13 and Xa21. The resistance R alleles were amplified using five functional markers (FMs) to generate amplicons of 217, 103, 179, 381 and 595 bp in IRBB66. Amplicons of 198, 107, 87, 391 and 467 bp corresponded to susceptible alleles in Taiwanese japonica rice cultivars. In backcross breeding programmes, the multiplex PCR assay was integrated into selection from a population using BB resistance donor IRBB66 crossed to rice cultivar ‘Tainung82’. Two plants with homozygosity for Xa4, xa5, Xa7, xa13 and Xa21 were selected from 1100 BC₂F₂ plants. In addition, the five BB resistance genes were also accurately identified in F₂ populations. This multiplex PCR method provides a rapid and efficient method for detecting various BB resistance genes, which will assist in pyramiding genes to improve durability of BB resistance in Taiwanese elite rice cultivars.     

Physiological Response of Multiple Contrasting Rice (Oryza sativa L.) Cultivars to Suboptimal Temperatures

The physiological response of multiple rice cultivars, eighteen initially and eight cultivars later on, to suboptimal temperatures (ST) conditions was investigated in laboratory and outdoor experimental conditions. Treatment with ST decreased growth in different extents according to the cultivar and affected the PSII performance, determined by chlorophyll fluorescence fast‐transient test, and stomatal conductance, regardless the experimental condition. Two groups of cultivars could be distinguished on the base of their growth and physiological parameters. The group of cultivars presenting higher growths displayed optimal JIP values, and higher instantaneous water use efficiency (WUE_i), due to a lower Gs under ST, unlike cultivars showing lower growth values, which presented worse JIP values and could not adjust their Gs and hence their WUE_i. In this work, we detected at least two cultivars with superior tolerance to ST than the cold tolerant referent Koshihikari. These cultivars could be used as parents or tolerance donors in breeding for new crop varieties. On other hand, positive and significant correlations between data obtained from laboratory and outdoor experiments suggest that laboratory measurements of most of the above mentioned parameters would be useful to predict the response of rice cultivars to ST outdoor.     

Deployment of cold tolerance loci from Oryza sativa ssp. Japonica cv. ‘Nipponbare’ in a high‐yielding Indica rice cultivar ‘93‐11’

Cold tolerance is a complex trait, and QTL pyramiding is required for rice breeding. In this study, a total of seven QTLs for cold tolerance in the Japonica rice variety ‘Nipponbare’ were identified in an F_2:3 population. A stably inherited major QTL, called qCTS11, was detected in the region adjacent to the centromere of chromosome 11. In a near‐isogenic line population, the QTL was further dissected into two linked loci, qCTS11.1 and qCTS11.2. Both of the homozygous alleles of qCTS11.1 and qCTS11.2 from ‘Nipponbare’ showed major positive effects on cold tolerance. Through pyramiding the linked QTLs in the cold‐sensitive Indica rice cultivar ‘93‐11’, we have developed a new elite, high‐yielding Indica variety with cold tolerance.     

Genetic dissection of vitamin E content in rice (Oryza sativa) grains using recombinant inbred lines derived from a cross between 'Zhenshan97B' and 'Nanyangzhan'

Vitamin E (VE) is an important nutritional trait in rice grains. In order to dissect the genetic basis underlying VE content, a recombinant inbred lines population derived from 'Zhenshan 97B' and 'Nanyangzhan' was used for quantitative trait locus (QTL) analysis. Totally, 29 QTLs for six VE traits were identified in 2 consecutive years. Among those, five QTLs repeatedly detected in two years formed a cluster on chromosome 2, which was responsible for all five VE isomers. OsγTMT, the gene encoding γ‐tocopherol methyltransferase in rice, was located to the same region and treated as the candidate gene. Sequence analysis of alleles from two parents revealed many polymorphisms, including 19 single nucleotide polymorphisms (SNPs) and two insert/deletions (Indels) in the promoter region, two nonsynonymous SNPs in exons, and 25 SNPs and an Indel in introns. Besides, a QTL for δ‐tocotrienol and two QTLs for α‐tocopherol were repeatedly detected on chromosome 5 and 8, respectively, all three regions carrying no homologous genes involved in VE biosynthesis. These results could be useful in development of rice lines displaying desirable VE content.     

水稻广亲和性的鉴定与研究

选用9个待测广亲和系，并以4个标准测验品种和7个光（温）敏核不育系为亲和性测验种，按NCII交配设计配制99个组合。根据亲本的籼稻分类地位和F1杂种结实率对待测广亲和系的广亲和性进行了鉴定与研究。标准测验品种检验表明，培矮64、CPSLO-17、02428具有广亲和性，轮回422、培C311、CY8543具有一定的广亲和性，皖恢9号、早轮     

Fine mapping of a day‐length‐sensitive dwarf gene in rice

Dwarf mutants are valuable and crucial resources for genetic research and crop breeding programme in rice. In this study, we identified a dwarf mutant derive from tissue culture, which exhibited a delayed heading date and dwarfism under long‐day growth conditions, suggesting the heading date of dwarf mutant was sensitive to day length. Based on 2000 F₂ mutant‐like individuals from the cross of the mutant and a Japonica var. ‘IRAT129’, the dwarf gene was finally narrowed into a 512‐kb region near the centromere on chromosome 9. According to the sequence analysis of a delimited region, 21 genes had base alternations either in promoters (15 SNPs) or in coding regions (6 InDels) among 73 annotated genes, and five genes were confirmed sequence alternations resulting from their expression mainly in the vegetative organs. Given to the RNAi plants of the five genes incapable to mimic dwarf and late heading date phenotype, the candidate gene remains to be identified by other genetic or molecular methods. Therefore, all these results give us informative foundation for the day‐length‐sensitive dwarf gene isolation.     

Sexual hybridization between Capsella bursa‐pastoris (L.) Medik (♀) and Camelina sativa (L.) Crantz (♂) (Brassicaceae)

The development of transgenic oilseed Camelina sativa (2n = 40) and the potential for hybridization with its weedy relative Capsella bursa‐pastoris (2n = 36) necessitates a careful evaluation of the reproductive compatibility between the species. Here, we conducted over 1800 crosses (emasculation and manual pollination) to examine the ability of 10 Canadian C. bursa‐pastoris (♀) accessions to hybridize with five accessions of C. sativa (♂). Seven hybrids were confirmed among 586 putative hybrids screened with species‐specific markers, indicating a hybridization rate of 1.5 hybrids per 10 000 ovules pollinated. All seven hybrids had intermediate DNA content compared to their parents, were morphologically distinct, had low (1.9%) pollen fertility and failed to produce selfed or backcrossed seed. Given the abundance of C. bursa‐pastoris along field margins, hybrids will likely be generated in the wild, but they will be unable to establish lineages unless fertility is restored. The large number of crosses and the diversity captured by the use of multiple accessions resulted in strong statistical power and a high degree of confidence in the estimated hybridization rate.     

Development of AS‐PCR marker based on a key mutation confirmed by resequencing of Wx‐mp in Milky Princess and its application in japonica soft rice (Oryza sativa L.) breeding

Soft rice with low amylose content (AC) ranging by 5–15% is a unique type with special eating and appearance quality and has become popular in the rice market. We resequenced the Wx‐mp, a key allele from Milky Princess, a Japanese low AC variety, and found that the +473 mutation in exon 4 is the key mutation in both Wx‐mp and its ancestor allele, Wx‐mq from Milky Queen. Based on this functional mutation, an allele‐specific PCR (AS‐PCR) marker was developed and proven in a breeding population derived from a cross between a Chinese late variety Nan Keng 46 (Wx‐mp/Wx‐mp) and an early line Ning 63121(Wx‐b/Wx‐b). Based on the marker‐aided selection by our newly developed AS‐PCR marker for Wx‐mp and the known ST10 marker for Stvb‐i, a total of 12 Wx‐mp homozygotes were selected from 198 F₂ progenies, and four of them were immune to rice stripe virus (RSV) with averagely 11.3 days earlier heading than Nan Keng 46 without significant change in grain yield.     

Genetics of Panicle Exsertion in Cold-Tolerant Rice (Oryza sativa)

Four cold-tolerant rice varieties, viz. ‘Khonorullo’, ‘Namyi’, ‘Abor B’ and ‘Meghalaya-1’ were crossed with two cold-susceptible ones, viz. ‘Pusa 33’ and ‘Subhadra’ (DR92), in all possible combinations to study the inheritance of gene(s) governing panicle exsertion and their allelic relationship among cold-tolerant varieties. F₁ hybrids of all the crosses showed complete panicle exsertion indicating dominance of this trait. Segregation pattern of panicle exsertion in F₂ and backcrosses show that all the four cold-tolerant varieties possessed a single dominant gene designated as Ctr-1. Absence of segregation for panicle exsertion in an F₂ generation obtained from intercrosses of cold-tolerant varieties suggests that the dominant genes in all the four cultivars are allelic.     

水稻耐低磷种质的筛选与鉴定

本研究对281份不同的水稻（Oryza sativa L.）品种（品系）进行了耐低磷筛选和鉴定。在大田种植条件下,通过水稻部分性状耐低磷系数（－P/+P）的变异范围、平均值以及它们间的相关性分析,认为相对籽粒产量、相对成熟期的地上部生物重、相对植株分蘖期的分蘖数是较好的耐低磷筛选和评价的指标。并提出以偏低含磷量大田初筛及     

Impact of Humidity on Temperature-Induced Grain Sterility in Rice (Oryza sativa L)

High temperature‐induced grain sterility in rice is becoming a serious problem in tropical rice‐growing ecosystems. We studied the mechanism of high temperature‐induced grain sterility of different rice (Oryza sativa L) cultivars at two relative humidity (RH) levels. Four varieties of Indica and Japonica rice were exposed to over 85 % RH and 60 % RH at 36/30 °C, 34/30 °C, 32/24 °C and 30/24 °C day/night air temperatures from late booting to maturity inside sunlit phytotrons. Increasing both air temperature and RH significantly increased spikelet sterility while high temperature‐induced sterility decreased significantly with decreasing RH. Neither Indica nor Japonica rice types were superior to the other in the response of their spikelets to increased air temperature and RH. Increased spikelet sterility was due to increased pollen grain sterility which reduced deposition of viable pollen grains on stigma. Reduction in sterility with decreased RH was more due to decreased spikelet temperature than to air temperature. Thus the impact of RH should be considered when interpreting the effect of high temperature on grain sterility. Spikelet fertility was curvilinearly related to spikelet temperature. Grain sterility increased when spikelet temperature increased over 30 °C while it became completely sterile at 36 °C. The ability of a variety to decrease its spikelet temperature with decreasing RH could be considered as avoidance while the variability in spikelet sterility among varieties at a given spikelet temperature could be considered as true tolerance.     

Genetic analysis and fine mapping of tms9, a novel thermosensitive genic male‐sterile gene in rice (Oryza sativa L.)

Two‐line hybrid rice as a novel hybrid breeding method has huge potential for yield increasing and utilization of intersubspecific heterosis, and it is of major significance for the food security of rice‐consuming populations. Zhu1S is a thermosensitive genic male‐sterile line of rice with low critical temperature and excellent combining ability, which has been widely exploited as a female parent in Chinese two‐line hybrid rice breeding. Here, genetic mapping in F₂ populations was used to show that its male sterility is inherited as a single recessive gene and that responsible gene (termed tms9) lies on the short arm of chromosome 2. A high‐resolution linkage analysis which was based on the Zhu1S/R173 F₂ population found that the thermosensitive genic male‐sterile gene tms9 of Zhu1S was fine mapped between insertion–deletion (Indel) markers Indel 37 and Indel 57, and the genetic distance from the tms9 to the two markers was 0.12 and 0.31 cM, respectively. The physical distance between the two markers was about 107.2 kb. Sequence annotation databases showed that the two Indel markers (Indel 37 and Indel 57) were located on two BAC clones (B1307A11 and P0027A02). There are sixteen open reading frames (ORF) present in this region. The results of this study are of great significance for further cloning tms9 and molecular marker–assisted selection.     

水稻条斑花叶突变体生态st(t) 的鉴定与遗传定位

利用EMS诱变育成优良籼型恢复系缙恢10号,从其后代中鉴定出一个白色条斑花叶突变体st(t),在三叶期开始表现白斑,拔节期白斑变为不规则线状,一直保持到成熟。突变体叶绿素含量明显下降,类胡萝卜素含量显著升高。透射电镜观察表明,突变体的绿色叶片部位与野生型相比,在细胞结构上无明显差异,叶绿体发育正常;突变体的白化部位细胞结构异常,质体内多含有积聚在一起的嗜锇小球,不能发育出正常叶绿体所具有的类囊体和基质片层结构。遗传分析表明该性状受一对隐性核基因调控,利用1 500株西农1A/st(t)的F₂隐性定位群体,最终把St(t)基因定位在第6染色体SSR标记RM19745和RM19762之间,遗传距离分别为0.07 cM和0.27 cM,根据9311基因组序列推测,两标记之间的物理距离约为345 kb。这为St(t)基因的图位克隆和分子标记辅助育种奠定了基础。     

Field‐grown Cotton Exhibits Seasonal Variation in Photosynthetic Heat Tolerance without Exposure to Heat‐stress or Water‐deficit Conditions

Thermotolerance acclimation of photosystem II to heat and drought is well documented, but studies demonstrating developmental impacts on heat tolerance in field‐grown plants are limited. Consequently, climatic variables, estimated canopy temperature, predawn leaf water potential (Ψ_PD), and the temperature responses of maximum quantum yield of photosystem II (F_v/F_m), variable fluorescence (F_v/F₀), quantum yield of electron transport (φ_Eο) and efficiency of PSI electron acceptor reduction (REο/ABS) were characterized for Gossypium hirsutum at three sample times during the growing season (21 June, 2 July and 18 July 2013) under well‐watered conditions. The temperature decreasing a given photosynthetic parameter 15% from the optimum is referred to as T₁₅ and served as a standardized measure of heat tolerance. Ambient and estimated canopy temperatures were well within the optimal range for cotton throughout the sample period, and leaves were verified well watered using Ψ_PD measurements. However, T₁₅ varied with sample date (highest on July 2 for all parameters), being 2 °C (F_v/F₀) to 5.5 °C (φ_Eο) higher on July 2 relative to June 21, despite optimal temperature conditions and predawn leaf water potential on all sample dates. These findings suggest that even under optimum temperature conditions and water availability, heat tolerance could be influenced by plant developmental stage.     

Reinventing heterosis phenomenon through deployment of alien introgression lines in rice (Oryza sativa L.)

Introgression Lines (ILs) carrying alien genomic segments in the homozygous state may or may not be able to contribute positively to the phenotype on account of replacement of cultivated genome segment, however, the genetic elite in heterozygous condition can be attributed to the presence of both recipient and donor genome complements. Therefore, overdominance or pseudo‐overdominance effect at the heterozygous loci is anticipated. Set of 318 ILs, carrying several genomic segments from “A” genome donor wild species, were used for developing test hybrids with CMS line PMS17A. Of these, three hybrids observed significant yield advantage (>25%) over recurrent parents and checks. Parental ILs of these hybrids viz., IL326, IL901 and IL951 carried 5.62%, 2.52% introgression from Oryza rufipogon and 6.71% from Oryza nivara, respectively. These ILs were also crossed with their recurrent parent to develop introgression line hybrids (ILHs) to observe the contribution of the alien segment(s), in the homozygous and heterozygous state. The traits studied have higher mean value when genomic segments from wild species were in the heterozygous state as compared to the homozygous state.     

水稻单叶独立转绿型黄化突变体grc2的鉴定与基因精细定位

转绿型叶色突变体是研究植物叶绿体分化与发育的基础材料。grc2是利用60Co-γ射线诱变籼型三系保持系T98B后获得的单叶独立转绿型黄化突变体。grc2植株上任一叶片刚抽出时为黄色,在生长10 d左右后变绿,具有单叶不依赖于植株特定发育阶段而独立转绿的特性。与野生型T98B相比,grc2黄化叶片的总叶绿素和叶绿素b含量显著降低,叶绿体滞留在黄化质体阶段,表明grc2可能在叶片早期发育中起关键作用。遗传分析表明,grc2受1对隐性核基因独立控制;利用源于grc2/Nipponbare的F2群体的960个突变单株,将grc2基因定位在STS标记S254与S258之间约31 kb的范围内,该区域含有5个未报道过的注释基因。这些结果为grc2的克隆及功能研究提供了重要信息。     

Genetic Analysis of Resistance to Brown Planthopper in Rice (Oryza sativa L.)

Seventeen rice cultivate resistant to brown planthoppers were genetically analyzed using the Bangladesh insect population. Seven cultivars were found to have a single dominant gene for resistance. These genes segregated independently of the recessive resistance gene bph-5. Tae dominant resistance gene of ‘Swarnalata’ was designated Bph-6. In ten cultivars, resistance is conferred by single recessive genes. In eight cultivars, the resistance genes are allelic to bph-5. However, the recessive genes o: two cultivars are non-allelic to bph-5. The recessive gene of T12 is designated bph-7.     

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。     

水稻内稃扭曲突变体palea distortion 1(pd1)的鉴定与基因定位

在对籼稻缙恢10进行EMS(ethyl methane sulfonate)处理后的群体中,发现一个花器官突变体,主要表现为内稃扭曲并呈现外稃化特征,浆片数目增加且呈现稃状特征,雄蕊数目减少至1～4个,部分雄蕊的花丝呈现浆片化特征,暂将其命名为水稻颖壳扭曲突变体palea distortion 1(pd1)。遗传分析表明该突变性状受一个隐形单基因控制。利用群体分离分析法(bulked segregation analysis,BSA),将PD1基因定位在第2染色体的RM13693和RM13936之间,遗传距离分别为3.25cM和3.90cM。该研究结果为PD1基因的图位克隆奠定了基础。