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.     

Correlation of Chlorophyll Meter Readings with Gas exchange and Chlorophyll Fluorescence in Flag Leaves of Rice (Oryza sativa L.) Plants

Abstract

The objective of this study was to establish the correlation of the chlorophyll meter (SPAD) readings with the contents of chlorophyll (Chl) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the gross photosynthetic rate (P_g), and the maximum quantum yield of photosystem II (PSII) (F_v/F_m) in flag leaves of rice (Oryza sativa L.) in ripening stage. The SPAD readings significantly correlated with the Chl content, the Rubisco content, P_g and F_v/F_m (R² = 0.848, 0.648, 0.671 and 0.712, respectively), which suggests that the SPAD meter has the potential to estimate the photosynthetic capacity of the flag leaves. However, both P_g and F_v/F_m had a stronger relationship with the Rubisco content than the SPAD readings, indicating that the PSII photochemical and CO₂ assimilation capacities are strongly influenced by the Rubisco content. Therefore, accurate calibration would be indispensable to obtain the physiological information from the SPAD readings of flag leaves.     

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.     

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。     

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.     

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.     

Effects of Exogenous Glycinebetaine on Growth and Ultrastructure of Salt-Stressed Rice Seedlings (Oryza sativa L.)

Abstract

The effects of exogenously applied glycinebetaine on the salt-stress-induced inhibition of growth and ultrastructural damages in rice seedlings were investigated. Glycinebetaine was not effective in alleviating the NaCl-induced inhibition of root growth and rather enhanced the NaCl-induced inhibition. However, it was found to alleviate the inhibition of shoot growth induced by NaCl stress. Concentrations of Na were higher in salt-stressed plants than in unstressed plants. Stressed plants receiving glycinebetaine had a significantly lower Na and higher K concentrations in the shoots than the plants grown without application of glycinebetaine. Salinity induced ultrastructural damages in leaf such as swelling of thylakoids, disintegration of grana stacking and intergranal lamellae and destruction of mitochondria (deficiency of cristae, swelling and vacuolation). Such damages were largely prevented by pretreatment with glycinebetaine resulting in greening of the plants. In roots, the epidermis, cortex and root cap were more sensitive to salt stress than the meristem and stele. The most frequently observed ultrastructural alteration due to NaCl salinity was the formation of many large vacuoles in the root tip and root cap cells. The number of mitochondria was increased and they were aggregated in the cytoplasm of the root tip and root cap cells by treatment with NaCl or NaCl plus glycinebetaine. Glycinebetaine could not prevent the NaCl-induced ultrastructural damages in root cells. The effects of glycinebetaine to mitigate the ultrastructural damages in the chloroplast and mitochondria induced by NaCl might be due to the production of many vacuoles in root cells which may act to store Na and decrease its accumulation in the shoot.     

Effects of Salinity Stress on the Seminal Root Tip Ultrastructures of Rice Seedlings (Oryza sativa L.)

Abstract

Growth and structural changes in the seminal root tip of rice seedlings (Oryza sativa L. cv. Nipponbare) in response to NaCl salinity were studied. Seedlings were grown in agar medium with 0, 0.1, 0.3, 1.0, 2.0 and 3.0% NaCl(agar culture), and in water with 0, 0.01, 0.03, 0.06 and 0.1% NaCl (water culture). Seedling growth was significantly suppressed by higher concentrations of NaCl. The effect of NaCl appeared faster in water culture than in agar culture. In both agar and water cultures, root growth was markedly suppressed over shoot growth. Under saline conditions, epidermis, cortex and root cap cells appear to be damaged to a greater extent than the meristem and stelar cells. The most notable ultrastructural change in response to salinity was the development and increment of vacuoles, which seem to provide a space for accumulation of excess ions. Many electron dense deposits were observed in the larger vacuoles of the epidermal and cortical cells. Under saline conditions, cell wall thickening of the epidermis, an increase in endoplasmic reticulum, myelin figures, less compact Golgi bodies and inhibited production of Golgi vesicles were also observed.     

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.     

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...     

Comparison of Susceptibility to Photoinhibition and Energy Partitioning of Absorbed Light in Photosystem II in Flag Leaves of Two Rice (Oryza sativa L.) Cultivars that Differ in Their Responses to Nitrogen-Deficiency

Abstract

Abstract: The energy partitioning in photosystem II (PSII) and the susceptibility to photoinhibition in PSII were investigated in flag leaves of two rice cultivars, Shirobeniya (a traditionaljaponica) and Akenohoshi (an improved japonica-indica intermediate) grown under standard-nitrogen (N) (SN) and low-N (LN) conditions. N-deficiency resulted in significant decreases in total dry weight, net photosynthetic rate (P_n), the energy flux via carboxylation (J_c), and content of ribulose-1,5-bisphosphate carboxylase /oxygenase (Rubisco) in flag leaves in the two cultivars, and these parameters of Shirobeniya were lower than those in Akenohoshi under the LN condition. In the two cultivars, the energy flux via alternative electron flow was significantly increased by N-deficiency, which was accompanied by enhanced activity of superoxide dismutase (SOD). Although under the LN condition no cultivar differences were found in J_a and SOD, ascorbate peroxidase activity in Shirobeniya was lower than that in Akenohoshi. N-deficiency resulted in more significant increases in the susceptibility to photoinhibition (the degree of decrease in maximum quantum yield of PSII), hydrogen peroxide (H₂O₂) content and malondialdehyde content after exposure to high irradiance in Shirobeniya than those in Akenohoshi. These results indicated that the increased susceptibility to photoinhibition in the LN plants of Shirobeniya was mainly due to oxidative damages to chloroplasts, resulting from lower carboxylation and H₂O₂-scavenging capacities. Therefore, both carboxylation and H₂O₂-scavenging capacities could be important factors in determining the cultivar difference in the productivity of rice under LN conditions.     

“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。     

The Promotive Effect of Brassinolide on Lamina Joint-Cell Elongation,Germination and Seedling Growth under Low-Temperature Stress in Rice (Oryza sativa L.)

Summary

The cell elongation-promoting activities of brassinolide (BR) and indoleacetic acid (IAA) in the lamina-inclination test using etiolated rice seedlings were compared at a low temperature. IAA promoted the lamina joint-cell elongation at 30°G, but the effect was markedly lowered with decreasing temperature and reached null at 15°C. On the other hand, BR and castasterone (CAS), a brassinosteroid, promoted this cell elongation even at 15°G. Homobrassinolide (HBR), a brassinosteroid, also promoted this cell elongation, although the activity was weak. The combination of BR and IAA synergistically promoted this cell elongation at a low temperature (15°G).

Immersion of the dry seeds in 2 X 10^-8 M BR for the first 24 h promoted the early growth of rice seedlings. This treatment also enhanced the germination rate and the growth after direct sowing in submerged paddy pots in a greenhouse or phytotron at a low temperature (15°C). Leaf spraying of BR on the rice seedlings at the 4th leaf stage increased plant height and the fresh weights of tops and roots even at a low temperature. BR sprayed on the completely expanded 4th leaves did not increase their blade length, but that sprayed on the expanding 5th and 6th leaves strikingly increased their blade length, and the effect tended to be stronger at a low temperature.

These results suggest that BR promotes cell elongation in young rice seedlings under low-temperature stress, although IAA had no such effect, and that BR may promote germination and the early growth of rice seedlings at a low temperature in direct sowing in the submerged paddy field and in the rice nursery.     

利用同工酶标记水稻亲本遗传差异及其在杂交水稻育种中的利用

利用淀粉凝胶和聚丙烯酰胺凝胶电泳显示的同工酶差异，分析17个杂交水稻亲本、3个新株型株系和22个光壳稻、爪哇稻品种的遗传差异，研究光壳稻和爪哇稻及其与温带粳稻之间的关系，探索同工酶标记水稻亲本遗传差异在杂交稻育种中利用的可能性。在本研究中，8个同工酶18个多态性位点至少在2个基因型间存在差异。每个具有多态性的酶谱带分别以1和0记录存在与否，由同工酶数据计算的Nei’s遗传距离创建聚类树状图。聚类分析结果表明，籼稻和粳稻容易被分开，普通粳稻与光壳稻、爪哇稻混合聚在一起；光壳稻与爪哇稻之间的遗传距离要比它们与普通粳稻之间的遗传距离小。利用光壳稻、爪哇稻选育不同生态群方向的恢复系和不育系，已配组育成了强优势的杂交稻组合。     

Enclosed stigma contributes to higher spikelet fertility for rice(Oryza sativa L.) subjected to heat stress

With global warming, rice plants may be subjected to heat stress more regularly during the heatsensitive flowering stage, causing spikelet sterility and grain yield loss.Stigma exsertion is considered to increase pollen reception and promote female reproductive success.The aim of this study was to investigate the role of stigma exsertion on spikelet fertility at high temperatures.Five rice cultivars(Liangyoupeijiu, Shanyou 63, Huanghuazhan, Nagina 22, and IR64) with differing degrees of stigma exsertion were cultivated and exposed to high temperature at anthesis.Heat-tolerant cultivars did not always show a high percentage of spikelets with exserted stigmas, and vice versa.Irrespective of the presence of more pollen grains on exserted stigmas, spikelets with exserted stigmas did not show greater spikelet fertility than spikelets with fewer exserted stigmas or hidden stigmas under heat stress.GA3 application augmented the percentage of spikelets with exserted stigmas;however, it did not increase spikelet fertility under heat stress.Spikelet fertility of whole panicles was negatively correlated with the percentage of spikelets with exserted stigmas, but positively with that with hidden stigmas.Viability of the hidden stigmas was less reduced than that of exserted stigmas under heat stress, suggesting that hidden stigmas have an advantage in maintaining viability.Heat stress delayed anther dehiscence and reduced the viabilities of both exserted stigmas and pollens, thereby causing low spikelet fertility.Together, these results suggest that high spikelet fertility does not depend on stigma exsertion and that enclosed stigma generally contributes to higher spikelet fertility and heat tolerance under high-temperature conditions during flowering in rice.     

水稻花粉发育过程及其分期

以籼稻品种IR36为材料,采用塑料半薄切片技术、新鲜花药整体观察及游离的雄性细胞观察方法,对水稻花粉发育全过程及其药壁组织的情况进行了系统的研究,初步将这一过程划分为8个时期,即小孢子母细胞形成期、小孢子母细胞减数分裂期、小孢子早期、小孢子中期、小孢子晚期、二胞花粉早期、二胞花粉晚期和成熟花粉期。并证实了花粉自然发育过程中不存在收缩期。在这一发育过程中,绒毡层出现3种明显不同的发育形态,这些形态特点对于判断花粉发育阶段具有重要的参考价值及辅助作用。     

Grain Protein Content of Interspecific Progenies Derivedfrom the Cross of African Rice (Oryza glaberrima Steud.) and Asian Rice (Oryza sativa L.)

Abstract

The protein contents of the grain of 50 interspecific progenies developed from the cross between WAB56-104, an Oryza sativa variety, and CG 14, an Oryza glaberrima line, were investigated. In contrast to the higher protein content of O. glaberrima than O. sativa on the average, the protein content of CG 14 was always lower than that of WAB56-104. However, judging from the average of three seasons, 72% of the interspecific progenies had a higher protein content than the mid-parent and 50% of them had a higher protein content than WAB56-104. Although the actual values of protein content of the interspecific progenies were significantly different among the seasons, a highly significant correlation was always observed in protein content between any two of the three seasons. Protein content therefore was considered character of each interspecific progeny though it was also affected by environment. A significant correlation was not observed between paddy yield and protein content in any season; several interspecific progenies showed higher protein content and paddy yield than the mid-parents. A low paddy yield is likely to be associated with high protein content through physiological regulation without a genetic linkage between the two traits. However, the results suggest that the transgressive segregation of protein content observed in the interspecific progenies is attributed not to this physiological regulation but to a certain mechanism to concentrate protein in grains with a genetic background.     

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.     

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。