The QTL Analysis of RuBisCO in Flag Leaves and Non-Structural Carbohydrates in Leaf Sheaths of Rice Using Chromosome Segment Substitution Lines and Backcross Progeny F2 Populations

Abstract

In rice (Oryza sativa L.), the maintenance of high photosynthetic rate of flag leaves and the carbon remobilization from leaf sheaths after heading is a critical physiological component affecting the yield. To clarify the genetic basis of RuBisCO content of the flag leaf, a major determinant of photosynthetic rate, and non-structural carbohydrate (NSC) concentration in the third leaf sheath at heading, we carried out quantitative trait loci (QTL) analysis with 39 Koshihikari/Kasalath chromosome segment substitution lines (CSSLs) and backcross progeny F₂ population derived from target CSSL holding the QTL/Koshihikari in the field. QTLs for RuBisCO content and NSC concentration at heading were detected between R2447-C1286 and R2447-R716 on chromosome 10, respectively, by comparing Koshihikari with four CSSLs for chromosome 10 (SL-229, -230, -231 and -232). The progeny QTL for RuBisCO content and for NSC concentration at heading qRCH-10 and qNSCLSH-10-1, respectively, were detected at similar marker intervals between RM8201 and RM5708. In addition, QTLs for RuBisCO content at 14 d after heading, qRCAH-10-1 and qRCAH-10-2, were detected in regions different from that of qRCH-10. No QTL for NSC concentration at 14 d after heading was detected between RM8201 and R716, the region analyzed in this study. The QTLs qRCH-10 and qRCAH-10-1 for RuBisCO content would have additive effects. These QTLs for RuBisCO content and NSC concentration newly found using CSSLs and their backcross progeny F₂ population should be useful for better understanding the genetic basis of source and temporary-sink functions in rice and for genetic improvement of Koshihikari in terms of their functions.     

Ammonia Emission from Leaves of Different Rice (Oryza sativa L.) Cultivars

Abstract

NH₃ emission from leaves of three rice (Oryza sativa L.) cultivars, Akenohoshi, Shirobeniya and Kasalath, was examined using a simple open chamber system.　In the three cultivars, NH₃ emission rate (AER) and NH₄⁺ content of leaves decreased with decreasing NH₄⁺ concentration in the root medium, but these values differed significantly with the cultivar. In the daytime, AER, NH₄⁺ content and glutamine synthetase (GS) activity in leaves changed similarly with maximum values around midday. Akenohoshi showed significantly lower AER and NH₄⁺ content but higher GS activity than Kasalath. The difference in AER among the rice cultivars may be related to the activity of GS involved in photorespiratory NH₃ recycling. Akenohoshi can be a breeding material useful for improving N recycling.     

几个籼稻品种的成熟胚愈伤组织植株再生体系的建立

组织培养技术在作物改良上的成功应用需要合适的植株再生体系。本试验以7个籼稻成熟胚为材料，通过优化激素配比，建立适合于水稻遗传转化的高频植株再生体系。研究结果表明，NB 2mg／L2，4-D适合于供试品种的成熟胚愈伤组织的诱导，诱导率达90％以上；在分化培养基中添加3．0-3．5mg／L KT，0．5～1．0mg／L NAA和5．0mg／L ABA的激素配比，明恢81、N175、航1号的最高分化率分别达72．7％、80．0％和78．0％；移栽成活95％以上。     

利用染色体片段置换系群体定位和分析水稻粒重和粒型QTL

[目的]挖掘水稻粒重和粒型相关性状QTL,对于解析水稻籽粒遗传机理具有重要作用.[方法]本研究以籼稻9311为受体、粳稻日本晴为供体构建的染色体片段置换系(Chromosome Segment Substitution Lines,CSSLs)群体为材料,在4个环境下对控制稻谷与糙米的粒重和粒型QTL进行了定位分析.[...     

Responses of Eighteen Rice (Oryza sativa L.) Cultivars to Temperature Tested Using Two Types of Growth Chambers

Abstract

Genetic variation in the growth response to temperature is a basis for developing adaptation measures to global warming, but evaluation of cultivars for the temperature responses may depend on other environmental factors such as light. In this study, we tested the growth responses of 18 diverse rice cultivars to constant day/night temperature of 25, 28, 31 and 34ºC in artificially-lit growth chambers (ALC) in Wagga Wagga (7.8 MJ m-² d-¹), and in naturally-lit chambers (NLC) in Yanco (25 and 28ºC and 13.4 MJ m-² d-¹; 31 and 34ºC and 11.5 MJ m-² d-¹), both in NSW, Australia. There was a significant interaction between temperature and chamber type for total shoot and panicle biomass; total shoot biomass was largest at 31ºC in ALC, and at 25 and 28ºC in NLC. From the average of all temperatures, the total shoot biomass declined by 29.5% in plants grown in ALC compared with those grown in NLC. Importantly, cultivar performance in ALC was similar to that in NLC at these temperatures, as evidenced by the highly significant correlation in total shoot biomass between ALC and NLC. Among 18 cultivars, IR64, IR72, N22, Vandana, Takanari and Koshihikari commonly produced a larger total shoot biomass under higher temperature conditions. Leaf area at earlier measurement date was highly correlated with the final total shoot biomass at the higher temperature more than specific leaf area.     

不同生态环境下水稻穗部性状QTL鉴定

【目的】发掘与产量相关的穗粒性状QTL对进一步克隆和利用高产基因具有重要意义。【方法】以超级粳稻龙稻5号和典型高产籼稻中优早8号杂交衍生的重组自交系群体为试材,在4种环境下对穗部性状进行比较和QTL分析。【结果】共检测到63个穗部性状QTL,分布于除第9染色体外的11条染色体上。在4个环境下分别检测到27、27、18和35个QTL。其中,16个QTL能在2个环境下被检测到,12个在3个以上环境下稳定表达,分别占QTL总数的25.40%和19.05%;第1、3、4和5染色体的多效QTL簇能在不同环境下稳定表达,对穗部性状具有明显的调控作用。【结论】第3染色体STS3.3-STS3.6区间的qSNP3、第4染色体RM5688-RM1359区间的qSNP4.1是2个新的稳定表达的多效性QTL簇。此外,上位性效应是调控穗部性状的重要组分。     

Identification of Casparian Bands in the Mesocotyl and Lower Internodes of Rice (Oryza sativa L.) Seedlings Using Fluorescence Microscopy

Abstract

The mesocotyl and lower internodes of seedlings are underground organs that play a possible role in water absorption and transport. The aim of this study is to reexamine and understand further the anatomy and morphology of mesocotyls and lower internodes in rice, especially in terms of the existence of Casparian bands. Anatomical structures of mesocotyl and lower internodes tissue of rice seedlings were observed by light and fluorescence microscopy. The rice mesocotyl had two central cylinders, one is large and the other small, in the cross section. Casparian bands were observed in both the endodermis and exodermis of the mesocotyl, and also in the endodermis of the first internode. Furthermore, some bundles with intermediate xylem differentiation were observed between the exarch and endarch arrangement in the first internodes. Casparian bands were identified in the radial walls of cells surrounding each of these bundles. The second internode is quite similar to that of upper internodes in adult plants with respect to internal structure; scattered vascular bundles were seen as in monocotyledons. In the second internode, Casparian bands were seen in the radial cell walls of the bundle sheaths in each vascular bundle. Unlike the mesocotyl, Casparian bands were not observed in the hypodermis of the first or second internodes. The results show that the histological features of the mesocotyl and lower internodes of rice seedlings widely differed. In addition, the present study provides anatomical evidence for the existence of Casparian bands in both the mesocotyl and lower internodes of rice seedlings.     

“三系”不育系微效恢复基因表现型研究

对胞质雄性不育系的3个育性指标:套袋自交结实率、黑染花粉率、嵌合颖花率,在4个不同育性水平的不育系的多个转育世代中的表现进行了比较。结果表明,3个指标均是微效恢复基因的遗传表现型,但嵌合颖花率因为易于考查、数据绝对值大、表现稳定性好而最适合作为微效恢复基因遗传分析的量化指标。该结论的重要意义在于量化的表现型是微效恢复基因遗传规律及进一步分子研究的基础。     

一份水稻OsWOX3B基因敲除突变体的鉴定

[目的]水稻OsWOX3B基因调控叶片形态和表皮毛发育,根据表型被命名为LSY1、DEP、NUDA和GLR1等.深入了解OsWOX3B基因对水稻发育调控的功能具有重要意义.[方法]利用CRISPR/Cas9基因编辑技术对籼稻品种R401的OsWOX3B进行基因敲除.对所获材料进行突变位点分析和表型分析,同时进行相关基因...     

利用染色体片段置换系定位水稻芽期耐冷性QTL

以籼稻品种9311为受体、粳稻品种日本晴为供体构建的95个染色体片段置换系为材料,在5℃低温条件下进行芽期耐冷性鉴定。结果表明,6个置换系低温处理后的成苗率与受体亲本9311有一定差异,其耐冷性略强于9311。利用代换作图法共鉴定出4个与芽期耐冷性相关的QTL,分别位于水稻第5和第7染色体上。其中qCTB-5-1、qCTB-5-2和qCTB-5-3分别被定位在第5染色体RM267与RM1237、RM2422与RM6054及RM3321与RM1054之间遗传距离分别为21.3cM、27.4cM和12.7cM的置换片段上;qCTB-7被定位在第7染色体RM11-RM2752区间遗传距离为6.8cM的置换片段上。     

Identification of QTLs for Plant Height and Its Components by Using Single Segment Substitution Lines in Rice （Oryza sativa）

QTLs for plant height and its components on the substituted segments of fifty-two single segment substitution lines （SSSLs） in rice were identified through t-test （P〈0.001） for comparison between each SSSL and recipient parent Huajingxian 74. On the 14 substituted segments, 24 QTLs were detected, 10 for plant height, 2 for panicle length, 4 for length of the first internode from the top, 5 for length of the second internode from the top and 3 for length of the third internode from the top, respectively. All these QTLs were distributed on nine rice chromosomes except chromosomes 5, 9 and 11. The additive effect ranged from -4.08 to 3.98 cm, and the additive effect percentages varied from -19.35% to 10.43%.     

Salinity-Induced Ultrastructural Alterations in Leaf Cells of Rice (Oryza sativa L.)

Summary

High concentrations of NaCl significantly reduced the fresh and dry weights and lengths of roots and shoots. NaCl exhibited a more rapid effect in water culture than in soil culture. In both water and solid cultures, root growth was suppressed more severely than shoot growth. Electron microscopic studies revealed that NaCl caused swelling of thylakoids, accumulation of starch grains and lipid droplets, distortion of grana stacking, increase in the size and number of plastoglobuli and vesiculation of cellular membrane. Mitochondria became deficient in cristae, swelled and the matrix appeared pale in salt-treated plants as compared with control plants. Disappearance of nucleolus and nuclear chromatin and destruction of vascular tissues were occasionally observed in salt-treated plants.     

Genotypic Variation of Stomatal Conductance in Relation to Stomatal Density and Length in Rice (Oryza sativa L.)

Abstract

Stomatal conductance (g_s) is an important trait responsible for the genotypic difference in gas diffusion for photosynthesis and transpiration in rice (Oryza sativa L.). We measured g_s, stomatal density and stomatal length (guard-cell length) at two weeks before heading for 64 accessions from a rice diversity research set of germplasm (RDRS) and for three high-yielding cultivars (HYC) under field conditions. Considerable variations in g_s, and stomatal length were observed among varieties in RDRS, and it was considered that RDRS covers the species diversity of the stomatal characteristics in rice. When it was compared among the varieties with similar plant earliness, g_s was higher in HYC than in most varieties of RDRS. Stomatal density did not correlate with g_s, and there was a negative correlation between stomatal density and stomatal length. However, noticeable variance existed in the latter relation, where HYC exhibited a higher stomatal density and slightly shorter stomatal length than RDRS. High g_s in HYC is attributable to their high stomatal density and moderate specific stomatal conductance (g_s / stomatal density) while the high-g_s varieties in RDRS tended to have a lower stomatal density and higher specific stomatal conductance. Stomatal length is related to specific stomatal conductance, but there are remarkable differences between these traits. Specific stomatal conductance in HYC has not reached the upper limit for their stomatal size, which raises a possibility of further improvement of HYC in g_s.     

Correlation between Chloroplast Ultrastructure and Chlorophyll Fluorescence Characteristics in the Leaves of Rice (Oryza sativa L.) Grown under Salinity

Abstract

The seedlings of Oryza sativa L. cv. Nipponbare grown by hydroponic culture for 3 wks were treated with 75, 100, 150 and 200 mM NaCl for 14, 14, 6 and 3 days, respectively, and examined for chloroplast ultrastructure in the region where chlorophyll fluorescence had been recorded. NaCl treatment decreased the ratio of variable to maximum chlorophyll fluorescence yield (F_v/F_m) and caused swelling of thylakoids. The swelling of thylakoids was quantified by the percentage of the length of swollen thylakoids to the total length of thylakoids. This value was increased with increasing NaCl concentration. Although F_v/F_m decreased at all concentrations of NaCl, the minimal fluorescence yield F₀ was not increased by the treatment with 75 or 100 mM NaCl. The percentage of the length of swelling was low at 75 and 100 mM NaCl. On the other hand, F₀ increased and the swelling of thylakoids was prominent with 150 and 200 mM NaCl treatment. These results suggest that the decrease in F_v/F_m due to the increase in F₀ under salt stress correlates with the ultrastructural damage. The decrease in F_v/F_m due to the increase in F₀ is expected to be useful as an indicator to evaluate the damage in chloroplasts, especially in thylakoid membranes, under salinity.     

外源磷处理对水稻籽粒植酸含量及相关代谢基因表达的影响

利用水稻穗离体培养方法,对不同外源磷浓度下水稻籽粒植酸等磷化物含量的差异及其植酸代谢相关功能基因在灌浆过程中的表达特征进行了探讨。结果表明,水稻籽粒中的植酸、无机磷和总磷含量随外源磷处理浓度上升呈增加趋势,但磷处理对千粒重和单位籽粒中植酸积累量的影响因磷浓度水平而异,高磷处理会导致水稻千粒重和籽粒植酸积累绝对量的显著降低;外源磷处理浓度的上升,不仅会引起水稻籽粒中锌和铁元素含量的显著降低,而且会导致锌、铁营养的生物有效性下降;外源磷处理对水稻籽粒植酸含量的影响与不同磷处理下RINO1基因的相对表达量之间存在较密切关系,中磷浓度(3P)处理会诱导RINO1基因的表达,但高磷浓度(12P)会抑制RINO1基因的表达,RINO1是外源磷浓度处理对水稻籽粒植酸合成代谢过程产生调控作用的一个重要功能基因位点,而IPK2基因与水稻在不同磷浓度处理下的植酸含量变化无直接联系。     

Characteristics of Gas Exchange and Chlorophyll Fluorescence during Senescence of Flag Leaf in Different Rice (Oryza sativa L.) Cultivars Grown under Nitrogen-Deficient Condition

Abstract

Effects of nitrogen (N) deficiency on photosynthetic gas exchange and photosystem II (PSII) photochemistry of flag leaves during grain-filling stage were investigated in six rice cultivars, Kasalath (a conventional indica), IR36 (an improved indica), Shirobeniya (a conventional japonica), Nipponbare (an improved japonica), Akenohoshi (an improved japonica-indica intermediate type) and BSI429 (an improved tropical japonica, a new plant type line) grown hydroponically in N-sufficient (NS) and N-deficient (ND) solution. From 3 to 24 days after heading (DAH), net photosynthetic rate (P_n), maximum quantum yield of photosystem II (PSII) (F_v/F_m), quantum yield of PSII electron transport ( Φ_psii), and contents of chlorophyll (Chl) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the flag leaves decreased, particularly under the ND condition in all six cultivars. A substantial difference was observed among the ND plants for the sustainability index (SI, the ratio of the value at 24 DAH to that at 3 DAH) of P_n, F_v/F_m, Φ_psii, Chl content and Rubisco content; SIs of those parameters of Akenohoshi, BSI429, Nipponbare and Shirobeniya were higher than those of IR36 and Kasalath. The SI of P_n showed significant positive correlations with those of F_v/F_m, Φ_psii, and the contents of Chl and Rubisco under the ND condition. It was concluded that the sustainability of photosynthesis in the flag leaves was mainly due to those of PSII photochemistry and electron transport, which was associated with the maintenance of Chl and Rubisco under the ND condition.     

Effects of Elevated Atmospheric Carbon Dioxide Concentration on Silica Deposition in Rice (Oryza sativa L.) Panicle

Abstract

The effects of elevated carbon dioxide concentration ([CO₂]) on silica deposition on husk epidermis of rice (Oryza sativa L. cv. Akitakomachi) during the flowering stage were investigated in this study. The study was motivated by the concept that the rice yield maybe affected by global warming as a result of elevated [CO₂] environment since sterility of rice is related to the panicle silica content that influences transpiration, and elevated [CO₂] could affect plant transpiration. Silica deposition analysis was focused on the flowering stage of the rice crop grown hydroponically under two [CO₂] conditions: 350 μmol mol-¹ (ambient) and 700 μmol mol-¹ (elevated). Silica deposition on the husk epidermis from three parts of the panicle at four flowering stages were examined using a scanning electron microscope (SEM) combined with an energy dispersive X-ray microanalyzer (EDX). The results demonstrated that elevated [CO₂] significantly suppressed silica deposition on the husk epidermis at the lower part of the panicle, and at the early flowering stage when 1/3 of the panicle emerged from the leaf sheath. In the transverse section analysis of the husk, silica deposition on the husk epidermis under elevated [CO₂] was less than that under ambient [CO₂] at the late flowering stage. The less silica deposition observed on the husks at the late flowering stage under elevated [CO₂] might be related to the suppressed transpiration from the panicle by elevated [CO₂] found in a previous study.     

Utilizing Chromosome Segment Substitution Lines (CSSLs) for Evaluation of Root Responses to Transient Moisture Stresses in Rice

Abstract

Drought and waterlogging that occur sequentially under field conditions are important abiotic stresses affecting plant growth and development. The ability to maintain the root system development during the contrasting moisture stresses may be one of the key traits for plant adaptation. This study aimed to identify the key root traits that contributed to the above ability by comparatively examining the effects of the two moisture stresses in succession on root system development. The chromosome segment substitution lines (CSSLs) from the crosses between the japonica rice cultivar Nipponbare and indica rice cultivar Kasalath were used for precise comparison of root system development. The rice seedlings were grown by hydroponics under a continuously well-aerated condition for 14 days (non-stressed), a drought condition for 7 days followed by an oxygen (O₂)-deficient (stagnant) condition for 7 days (drought-to-stagnant, D-S), or a stagnant condition for 7 days followed by drought condition for 7 days (stagnant-to-drought, S-D). CSSL43 and 47 did not show any significant differences in growth from Nipponbare under the non-stressed condition, but exhibited greater lateral root production under the stresses. Lateral root production was most closely related to faster seminal root elongation mediated by higher aerenchyma formation in the D-S condition, and to more branching of lateral roots on the seminal root axis in the S-D condition. The D-S condition severely affected lateral root production due to reduced seminal root elongation and aerenchyma formation. These results confirmed the fact that those root traits previously identified using different cultivars greatly contribute to plant adaptation. Oxygen deficiency preceded by drought (D-S) was more stressful to roots than drought preceded by O₂ deficiency (S-D), because drought reduced root aerenchyma formation during the subsequent stagnant condition.     

Rapid Swelling of Pollen Grains in Response to Floret Opening Unfolds Anther Locules in Rice (Oryza sativa L.)

Abstract

The changes in pollen grain diameter were examined in relation to the floret opening and anther dehiscence in rice (Oryza sativa L.). In the first experiment, the florets were artificially opened by excising the top of the glumes at various times before the expected natural flowering time. Pollen diameter increased rapidly in the artificially opened florets, but slowly in the controls in which the glumes were left intact. The time of anther dehiscence coincided well with the time when pollen grains reached their maximum diameter. In addition, the pollen grain diameter was significantly correlated with the percentage of dehisced anthers in the florets artificially opened at various times. These results indicate that floret opening induces the swelling of pollen grains and that the swelling is an important event for anther dehiscence. In the second experiment, anther segments with pollen left remaining intact and those with pollen removed were immersed in water. The septa in the anther segments without pollen failed to rupture in water, whereas 70% of the septa in the anther segments with pollen left remaining ruptured, indicating that the septa can be ruptured by the swelling pressure of pollen grains resulting in unfolding of anther locules. From these findings, we infer that the rapid swelling of pollen grains in response to floret opening is a driving force to rupture the septum and unfold the locules.     

Gene Expression of Enzymes for Starch and Sucrose Metabolism and Transport in Leaf Sheaths of Rice (Oryza sativa L.) during the Heading Period in Relation to the Sink to Source Transition

Abstract

Leaf sheaths of rice plants are known to temporarily accumulate starch prior to heading, which is subsequently remobilized and transported into the panicle after heading. We investigated the time course for both carbohydrate content and steady state mRNA levels of enzymes related to starch and sucrose metabolism in the rice leaf sheath (Oryza sativa L. cv. Nipponbare). Leaf sheaths from the second leaf below the flag leaf accumulated high levels of starch before heading but they rapidly decreased after heading. In contrast, the flag leaf sheath did not accumulate as much starch. In the second leaf sheath, the mRNA levels of enzymes involved in starch synthesis, ADP glucose pyrophosphorylase (EC 2. 7. 7. 27), soluble starch synthase (EC 2. 4. 1.21) and branching enzyme (EC 2. 4. 1. 18) were high before heading, which coincided with rapid accumulation of starch. The mRNA levels of sucrose synthesis enzymes, cytosolic FBPase (EC 3. 1. 3. 11) and sucrose phosphate synthase (EC 2. 4. 1. 14), and the sucrose transporter (OsSUTI) increased at the time of heading, which was largely coincident with a decrease in the mRNA levels of starch synthesis enzymes. In the flag leaf sheaths, changes in mRNA levels of starch synthesis enzymes were not pronounced, however mRNA levels of sucrose synthesis enzymes and the sucrose transporter showed a clear increase throughout the heading period. The different characteristics observed between the two leaf sheaths will be discussed in relation to the sink to source transition.