外源调节剂对淹涝水稻幼苗株高及碳水化合物消耗的影响

为研究淹涝条件下水稻幼苗株高及碳水化合物消耗对不同外源生长调节剂的响应,本试验选用籼型常规稻IR64和导入耐淹涝基因Sub1的IR64-Sub1为试验材料,秧龄20 d时喷施1-氨基环丙烷-1-羧酸(ACC)、多效唑(PB)、赤霉素(GA)3种外源生长调节剂,以喷施清水为对照(CK).喷施处理2 d后进行0、4、8、12、16 d没顶淹涝胁迫,淹涝胁迫结束后常温恢复7 d,取样分析不同外源生长调节剂对水稻成活率、株高、叶绿素降解及恢复、地上部非结构性碳水化合物(NSC)消耗的影响.结果表明:淹涝导致水稻幼苗株高显著增长,叶片SPAD值快速下降,叶片可溶性糖迅速消耗,但耐淹涝品种IR64-Sub1淹水前茎鞘NSC含量明显高于IR64,淹涝中NSC消耗速率低于IR64,淹水结束后地上部淀粉含量高于IR64.外源PB处理显著抑制水稻幼苗株高增长、叶绿素降解及NSC消耗,提高存活率,且对IR64-Sub1效果更为显著.外源GA处理水稻幼苗叶绿素降解、株高增长和NSC消耗最快,植株恢复能力最低,耐淹涝能力最差,但与IR64相比,GA对IR64-Sub1淹涝耐性的抑制明显减弱.外源ACC促进伸长效果明显低于外源GA处理.淹水前喷施PB可有效抑制植株水下伸长,延缓叶绿素降解,减缓NSC消耗,保留更多NSC,为淹水胁迫解除后水稻快速恢复提供有利条件,这对于易涝地区减轻涝渍危害具有重要意义.     

A major locus for submergence tolerance mapped on rice chromosome 9

Submergence stress is a widespread problem in rice-growing environments where drainage is impeded. A few cultivars can tolerate more than 10 days of submergence, but the genes conferring this tolerance have not been identified. We used randon-amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers to map submergence tolerance in 169 F₂ plants and the resulting F₃ families of a cross between a tolerant indica rice line, IR40931-26, and a susceptible japonica line, PI543851. IR40931-26 inherited strong submergence tolerance from the unimproved cultivar FR13A. Eight-day old F₃ seedlings were submerged for 14–16 days in 55-cm deep tanks, and tolerance was scored after 7 days recovery on a scale of 1 (tolerant) to 9 (susceptible). The tolerant and susceptible parents scored 1.5 and 8.4, respectively, and the F₃ means ranged from 1.6 to 8.9. Two bulks were formed with DNA from F₂ plants corresponding to the nine most tolerant and the nine most susceptible F₃ families. Of 624 RAPD primers used to screen the bulks, five produced bands associated with either tolerance or susceptibility. These markers were mapped to a region of chromosome 9 by linkage to RFLP markers. A submergence tolerance quantitative trait locus (QTL), here designatedSub1, was located ca. 4 cM from the RFLP marker C1232 and accounted for 69% of the phenotypic variance for the trait.     

Molecular mechanisms and future improvement of submergence tolerance in rice

Molecular Breeding - Low temperature (LT) is a key environmental stress affecting the growth, development, yield, and quality of wheat (Triticum aestivum L.). To better understand the genetic basis...     

Metabolic changes in rice seedlings with different submergence tolerance after desubmergence

When flash flood intolerant rice cultivars are submerged, they show greater morphological changes such as elongation and chlorosis than tolerant cultivars. These morphological responses are caused by ethylene produced during submergence, however, a visible damage of intolerant cultivars is markedly developing after desubmergence rather than during submergence, which is probably due to oxidative damage. We studied the effect of ethylene produced during submergence on antioxidant content and oxidative damage after desubmergence. When rice (Oryza sativa) was submerged for 8 days, both tolerant cultivar (BKNFR) and intolerant cultivars (Mashuri and IR42) showed a decrease in ascorbate concentration during submergence. After 3 days of desubmergence, the tolerant cultivar showed a rapid recovery of total ascorbate and ascorbic acid, whereas intolerant cultivars showed a slow recovery of them, an increase in malondialdehyde formation, and low survival rate (about 30%). However, applying 200 mg l⁻¹ of AgNO₃ as an ethylene antagonist to intolerant cultivars suppressed the decrease in ascorbate and the increase in malondialdehyde formation after desubmergence, and improved survival rate to about 60%. Ascorbic acid supply to leaf discs from submerged IR42 suppressed increase in malondialdehyde formation by incubation under the light for 24 h. In addition, strong negative correlations were observed between malondialdehyde formation with ascorbate concentration (r=−0.93) and with percentage of survival (r=−0.98). Our results indicate that the accumulated ethylene during submergence adversely affected antioxidant mechanism in intolerant rice cultivars after desubmergence, and ascorbic acid was an important antioxidant in vivo for the recovery of submerged rice seedlings.     

Diallel analysis of submergence tolerance in rice,Oryza sativa L.

Summary The genetics of submergence tolerance in rice was studied in a 10 × 10 half-diallel cross set involving 10 lowland rice varieties, four of which were tolerant (FR13A, FR43B, Kurkaruppan, and Goda Heenati) and the remaining six were nontolerant (RD19, IR42, IR17494-32-1, IR19672-24-3, Jagannath, and CR1009). Estimates of genetic parameters following Hayman's method showed significant additive and nonadditive gene action and the latter appeared to be solely due to dominance. Narrow sense heritability (0.70) indicated that additive gene effects were more important in the inheritance of the trait. Tolerance was dominant over nontolerance and the average dominance was within the range of incomplete dominance. Dominant alleles were more concentrated in the three tolerant parents, FR13A, Kurkaruppan, and FR43B in that order. Wr/Vr graphic analysis suggested the involvement of both major and minor genes. Combining ability analysis by Griffing's method also indicated significance of both additive and nonadditive effects, and the former appeared to be more important than the latter. The hybrids involving FR13A with RD19, IR42, and IR17494-32-1, and those of Kurkaruppan with RD19 and CR1009 appeared to be promising for incorporating an adequate level of tolerance to submergence into lowland rice cultivars.     

The key regulator of submergence tolerance,SUB1A,promotes photosynthetic and metabolic recovery from submergence damage in rice leaves

The submergence‐tolerance regulator, SUBMERGENCE1A (SUB1A), of rice (Oryza sativa L.) modulates gene regulation, metabolism and elongation growth during submergence. Its benefits continue during desubmergence through protection from reactive oxygen species and dehydration, but there is limited understanding of SUB1A's role in physiological recovery from the stress. Here, we investigated the contribution of SUB1A to desubmergence recovery using the two near‐isogenic lines, submergence‐sensitive M202 and tolerant M202(Sub1). No visible damage was detected in the two genotypes after 3 d of submergence, but the sublethal stress differentially altered photosynthetic parameters and accumulation of energy reserves. Submergence inhibited photosystem II photochemistry and stimulated breakdown of protein and accumulation of several amino acids in both genotypes at similar levels. Upon desubmergence, however, more rapid return to homeostasis of these factors was observed in M202(Sub1). Submergence considerably restrained non‐photochemical quenching (NPQ) in M202, whereas the value was unaltered in M202(Sub1) during the stress. Upon reaeration, submerged plants encounter sudden exposure to higher light. A greater capability for NPQ‐mediated photoprotection can benefit the rapid recovery of photosynthetic performance and energy reserve metabolism in M202(Sub1). Our findings illuminate the significant role of SUB1A in active physiological recovery upon desubmergence, a component of enhanced tolerance to submergence.     

Mapping QTLs for submergence tolerance in rice by AFLP analysis and selective genotyping

By combining the amplified fragment length polymorphism (AFLP) technique with selective genotyping, we constructed a linkage map for rice and assigned each linkage group to a corresponding chromosome. The AFLP map, consisting of 202 AFLP markers, was generated from 74 recombinant inbred lines (RIL) which were selected from both extremes of the population (250 lines) with respect to the response to complete submergence. Map length was 1756 cM, with an average interval size of 8.5 cM. To assign linkage groups to chromosomes, we used 50 previously mapped AFLP markers as anchor markers distributed over the 12 chromosomes. Other AFLP markers were then assigned to specific chromosomes based on their linkage to anchor markers. This AFLP map is equivalent to the RFLP/AFLP map constructed previously as the anchors were in the same order in both maps. Furthermore, tests with two restriction fragment length polymorphism (RFLP) markers and two sequence-tagged site (STS) markers showed that they mapped in the expected positions. Using this AFLP map, a major gene for submergence tolerance was localized on chromosome 9. Quantitative trait loci (QTL) associated with submergence tolerance were detected on chromosomes 6, 7, 11, and 12. We conclude that the combination of AFLP mapping and selective genotyping provides a much faster and easier approach to QTL identification than the use of RFLP markers. Received: 20 December 1996 / Accepted: 21 January 1997     

Effect of nutrient application and water turbidity on submergence tolerance of rice (Oryza sativa)

Flooding imposes severe selection pressure on plants, principally because excess water in their surroundings deprives the plants from certain basic resources such as oxygen, carbon dioxide and light for photosynthesis. Here, the effects of nitrogen and phosphorus application and their application time, on four Indica rice cultivars and their tolerance to submergence under turbid and clear flood water was studied. Submergence tolerance on metabolic changes, photosynthetic (Pn) rate and ethylene accumulation were evaluated. Submergence substantially reduced survival, chlorophyll, soluble sugars, Pn rate across cultivars with drastic reduction in IR‐20. Sub1 cultivars showed greater Pn rate as compared to sensitive cultivar under submergence because of better protection of chlorophyll, more stomatal conductance leading to higher survival. Turbid water resulted in lower under‐water photosynthesis because of poor light transmission, chlorophyll retention and silt deposition and ultimately poor survival. Foliar spray of N after desubmergence along with basal P improved the chlorophyll, soluble sugars and Pn rate and extenuated ethylene accumulation and shoot elongation resulting in significantly higher survival. To the best of our knowledge, this is the first time that such a study has been performed. Crop establishment could therefore be enhanced in areas where untimely flooding is anticipated by applying basal P and foliar spray of post‐submergence N.     

Identifying novel QTLs for submergence tolerance in rice cultivars IR72 and Madabaru

Short-term submergence is a recurring problem in many rice production areas. The SUB1 gene, derived from the tolerant variety FR13A, has been transferred to a number of widely grown varieties, allowing them to withstand complete submergence for up to 2 weeks. However, in areas where longer-term submergence occurs, improved varieties having higher tolerance levels are needed. To search for novel quantitative trait loci (QTLs) from other donors, an F_2:3 population between IR72 and Madabaru, both moderately tolerant varieties, was investigated. After a repeated phenotyping of 466 families under submergence stress, a subset of 80 families selected from the two extreme phenotypic tails was used for the QTL analysis. Phenotypic data showed transgressive segregation, with several families having an even higher survival rate than the FR13A-derived tolerant check (IR40931). Four QTLs were identified on chromosomes 1, 2, 9, and 12; the largest QTL on chromosome 1 had a LOD score of 11.2 and R ² of 52.3%. A QTL mapping to the SUB1 region on chromosome 9, with a LOD score of 3.6 and R ² of 18.6%, had the tolerant allele from Madabaru, while the other three QTLs had tolerant alleles from IR72. The identification of three non-SUB1 QTLs from IR72 suggests that an alternative pathway may be present in this variety that is independent of the ethylene-dependent pathway mediated by the SUB1A gene. These novel QTLs can be combined with SUB1 using marker assisted backcrossing in an effort to enhance the level of submergence tolerance for flood-prone areas.     

The biophysical basis of elongation growth in internodes of deepwater rice

Partial submergence induces rapid internodal elongation in deepwater rice (Oryza sativa L., cv Habiganj Aman II). We measured in vivo extensibility, tissue tension, hydraulic conductance and osmotic potential in the region of cell elongation in the uppermost internode. The in vivo extensibility of the internode, measured by stretching of living tissue with a custom-made constant stress extensiometer, rose rapidly following submergence of the plant. Both the elastic (E_el) and plastic (E_pl) extensibility increased when growth of the internode was induced. The submerged internode displayed tissue tension (elastic outward bending of longitudinally split internode sections); in air-grown control internodes, no such bending occurred. The hydraulic conductance, estimated from the kinetics of tissue shrinkage in 0.5 molar mannitol and subsequent swelling in distilled water, was not changed by submergence. The osmotic potential, measured with a dew-point hygrometer using frozen-thawed tissue, was only 18% less negative in the submerged internode than in the air-grown control. This indicates that osmoregulation takes place in rapidly elongating rice internodes. We suggest that the rapid expansion of the newly formed internodal cells of submerged plants is controlled by the yielding properties (E_pl) of the cell walls. Experiments with excised stem sections indicate that gibberellin is involved in increasing the E_pl of the elongating cell walls.     

长江下游不同类型水稻分蘖期耐淹能力比较

在分蘖期对长江下游稻作区主栽的9个水稻品种进行大田模拟没顶淹涝处理,研究淹涝胁迫对水稻植株农艺性状、生理指标和产量性状的影响,比较分析了常规粳稻、杂交籼稻和杂交粳稻对淹水胁迫环境适应性的差异.结果表明:淹水胁迫4d后,水稻株高及顶部全展3片叶长均比对照有不同程度的增加,伸长程度为杂交粳稻＞杂交籼稻＞常规粳稻.杂交粳稻的茎蘖数、绿叶数和地上部干质量损失率分别为18.0％、41.4％、13.2％,显著小于常规粳稻;杂交籼稻则介于杂交粳稻和常规粳稻之间,且整株死亡率显著低于常规粳稻.常规粳稻叶片中丙二醛(MDA)含量比对照增加1.91 μmol·g-1 FM,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性明显降低;杂交粳稻和杂交籼稻MDA含量分别降低2.32和2.10 μmol·g-1 FM,SOD和CAT活性显著提高.不同类型品种的减产程度差异显著,常规粳稻的产量损失率达到38.5％,显著高于杂交粳稻和杂交籼稻,杂交粳稻产量损失率仅为4.1％.长江下游水稻分蘖期的耐淹涝能力为杂交粳稻强于杂交籼稻,常规粳稻的耐淹能力最低.     

Biological activity of pestalotins on the elongation growth of rice seedlings

The effect of (–)-pestalotin, (+)-pestalotin and (+)-epipestalotinon the elongation growth of rice seedlings was studied. Thesecompounds did not influence elongation when given alone, although(–)-pestalotin and (+)-epipestalotin synergistically enhanced GA₃-induced elongation. (Received May 20, 1977; )     

Combining drought and submergence tolerance in rice: marker-assisted breeding and QTL combination effects

TDK1 is a popular rice variety from the Lao PDR. Originally developed for irrigated conditions, this variety suffers a high decline in yield under drought conditions. Studies have identified three quantitative trait loci (QTLs) for grain yield under drought conditions, qDTY _3.1 , qDTY _6.1 , and qDTY _6.2 , that show a high effect in the background of this variety. We report here the pyramiding of these three QTLs with SUB1 that provides 2–3 weeks of tolerance to complete submergence, with the aim to develop drought- and submergence-tolerant near-isogenic lines (NILs) of TDK1. We used a tandem approach that combined marker-assisted backcross breeding with phenotypic selection to develop NILs with high yield under drought stress and non-stress conditions and preferred grain quality. The effect of different QTL combinations on yield and yield-related traits under drought stress and non-stress conditions is also reported. Our results show qDTY _3.1 to be the largest and most consistent QTL affecting yield under drought conditions, followed by qDTY _6.1 and qDTY _6.2 , respectively. QTL class analysis also showed that lines with a combination of qDTY _3.1 and qDTY _6.1 consistently showed a higher tolerance to drought than those in which one of these QTLs was missing. In countries such as Lao PDR, where large areas under rice cultivation suffer vegetative-stage submergence and reproductive-stage drought, these lines could ensure yield stability. These lines can also serve as valuable genetic material to be used for further breeding of high-yielding, drought- and submergence-tolerant varieties in local breeding programs.     

A high-resolution linkage map of the vicinity of the rice submergence tolerance locus Sub1

Resistance to submergence stress is an important breeding objective in areas where rice cultivars are subjected to complete inundation for a week or more. The present study was conducted to develop a high-resolution map of the region surrounding the submergence tolerance gene Sub1 in rice, which derives from the Indian cultivar FR13A. Submergence screening of 8-day-old plants of F₃ families kept for 14 days submerged in 60 cm of water allowed an accurate classification of Sub1 phenotypes. Bulked segregant analysis was used to identify AFLP markers linked to Sub1. A population of 2950 F₂ plants segregating for Sub1 was screened with two RFLP markers flanking the Sub1 locus, 2.4 and 4.9 cM away. Submergence tolerance was measured in the recombinant plants, and AFLP markers closely linked to Sub1 were mapped. Two AFLP markers cosegregated with Sub1 in this large population, and other markers were localized within 0.2 cM of Sub1. The high-resolution map should serve as the basis for map-based cloning of this important locus, as it will permit the identification of BAC clones spanning the region. Received: 15 December 1999 / Accepted: 18 February 2000     

Evolutionary analysis of the Sub1 gene cluster that confers submergence tolerance to domesticated rice

BACKGROUND AND AIMS: Tolerance of complete submergence is recognized in a small number of accessions of domesticated Asian rice (Oryza sativa) and can be conferred by the Sub1A-1 gene of the polygenic Submergence-1 (Sub1) locus. In all O. sativa varieties, the Sub1 locus encodes the ethylene-responsive factor (ERF) genes Sub1B and Sub1C. A third paralogous ERF gene, Sub1A, is limited to a subset of indica accessions. It is thought that O. sativa was domesticated from the gene pools of the wild perennial species O. rufipogon Griff. and/or the annual species O. nivara Sharma et Shastry. The aim of this study was to evaluate the orthologues of the Sub1 locus in the closest relatives of O. sativa to provide insight into the origin of the gene and allelic variation of the Sub1 locus. METHODS: Orthologues of the Sub1 genes were isolated from O. rufipogon and O. nivara by use of oligonucleotide primers corresponding to the most highly conserved regions of the Sub1 genes of domesticated rice. The phylogenetic relatedness of Sub1 genes of O. sativa and its wild relatives was evaluated. KEY RESULTS AND CONCLUSIONS: Both O. rufipogon and O. nivara possess two Sub1 gene orthologues with strong sequence identity to the Sub1B and Sub1C alleles of cultivated rice. The phylogeny of the Sub1 genes of the domesticated and wild rice suggests that Sub1A arose from duplication of Sub1B. Variation in Sub1B alleles is correlated with the absence or presence of Sub1A. Together, the results indicate that genetic variation at the Sub1 locus is due to gene duplication and divergence that have occurred both prior to and after rice domestication.     

Submergence tolerance of rice – A new glasshouse method for the experimental submergence of plants

A new method is described for evaluation of submergence tolerance of rice ( Oryza sativa L.) plants. Responses of a range of cultivars corresponded with known differences in field performance. The method 1) allows fast and effective determination of submergence tolerance, 2) allows screening of many plants in a small glasshouse area, 3) provides for recovery of superior plants for seed collection, 4) allows manipulation of many environmental variables to mimic the natural submergence environment, and 5) uses simple, inexpensive, readily available equipment. Physiological studies performed with this method gave results similar to those obtained from field studies and showed that submergence tolerance increased in older plants; it decreased with increasing depth, increasing temperature and with high or low light levels. The system is ideal for the rapid evaluation of rice germplasm under controlled conditions and physiological studies on the mechanism of rice submergence tolerance.     

Studies on the growth of rice tissue under submerged condition. I. The effect of 2,4-dichlorophenoxyacetic acid on excised segments of coleoptiles under submergence

Summary The work presented deals with the fact that rice coleoptiles elongate more rapidly and more extensively under water than in air.Coleoptile segments of rice were cultured under submerged condition as well as under floating condition. On application of 2,4-D a sharp and significant increase in growth in elongation was recorded.At higher concentrations e. g., 100 and 10 p. p. m. the growth rate was higher in floating segments of coleoptiles. But at lower concentrations, including control, the growth rate was higher in submerged ones, which apparently indicates that the optimum concentration of 2,4-D for growth of rice coleoptile is shifted with shifting of oxygen tension. Three different mutually opposing factors namely, lowered auxin destruction under submergence, concentration of auxin in the plant tissue and lowered aerobic respiration have been stated to be responsible for growth of rice tissue under water.At the end we offer our sincere thanks to Dr. P. K.Sen, Khaira Professor and Head of the Department of Agriculture, University of Calcutta for granting all facilities to complete this investigation.