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.     

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.     

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.     

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.     

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.     

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.     

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.     

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。     

水肥一体化条件下施氮量对小麦旗叶叶绿素荧光特性及产量的影响

为探讨水肥一体化条件下施氮量对小麦旗叶叶绿素荧光特性及籽粒产量的影响,以小麦品种济麦22为试验材料,设每公顷施纯氮0（N0）、150（N1）、180（N2）、210（N3）和240 kg（N4）五个处理,通过大田试验分析了不同处理间小麦旗叶PSⅡ 最大光化学量子效率（F_v/F_m）、PSⅡ实际光化学效率（Φ_PSⅡ）、叶绿素相对含量、籽粒灌浆速率及产量的差异。结果表明,花后14~35 d,小麦旗叶F_v/F_m在N3和N4处理间无显著差异,但二处理显著高于其他处理。花后0~35 d,N3处理的旗叶Φ_PSⅡ最高,施氮量再增加至N4时,Φ_PSⅡ无显著变化。花后14~28 d,N3和N4处理的旗叶叶绿素相对含量显著高于其他处理;花后35 d,N3处理显著高于N4处理。粒重和灌浆速率在灌浆前期均随施氮量增加而降低;灌浆后期N3与N4处理间粒重和灌浆速率无显著差异,但二处理显著高于其他处理。籽粒产量在N3处理下最高;在施氮量增加至N4时,籽粒产量不再增加,且氮肥生产效率和氮肥农学效率分别下降了14.6%和24.0%。在本试验条件下,210 kg·hm^-2是兼顾高产和高效的最佳施氮量。     

Effect of Leaf Phosphorus and Potassium Concentration on Chlorophyll Meter Reading in Rice

Abstract

Chlorophyll meter (SPAD) is a convenient tool to estimate leaf nitrogen (N) concentration of rice plants. There is no information on the effects of leaf phosphorus (P) and potassium (K) concentration on SPAD readings and on the relationship between SPAD values and leaf N concentration in the literature. In 1996 dry season, cv IR72 was grown at the International Rice Research Institute (IRRI) and the Philippine Rice Research Institute (PhilRice) under various N, P and K fertilizer combinations. SPAD measurements were made on the topmost fully expanded leaves at mid-tillering and panicle initiation. The leaves were then detached, dried and analyzed for N, P and K. The SPAD values were highly correlated with leaf N concentration (r = 0.93 to 0.96). Fertilizer-K application did not affect SPAD values, leaf N concentration, or the relationship between the two. Phosphorus deficiency reduced leaf N concentration at mid-tillering, but increased leaf N concentration at panicle initiation when the same amount of N was applied. The SPAD values were 1 to 2 units greater for zero-P plants than P-treated plants at a given leaf N concentration at mid-tillering. At panicle initiation, the relationship between SPAD values and leaf N concentration was not significantly affected by leaf P status. These results suggest that a different regression equation between SPAD values and leaf N concentration should be used to estimate leaf N concentration of P-deficient and P-sufficient rice leaves at vegetative stage using a SPAD.     

Leaf Growth,Gas Exchange and Chlorophyll Fluorescence Parameters in Response to Different Water Deficits in Wheat Cultivars

Abstract

We investigated responses of wet climate (CY17) and dry climate (XN889) Trititcum aestivum L cultivars under 85, 55 and 25% field water capacity (FC). Less decrease in grain yield, relative water content, growth, gas exchange and chlorophyll fluorescence parameters indicated that XN889 was more drought-tolerant than CY17. At 55%FC, CY17 showed a lower net photosynthetic rate (Pn) than XN889 mainly due to stomatal closure. Stomatal closure was also observed in XN889, but its Pn was higher at 55%FC than at 85%FC. The higher Pn in XN889 may be associated with a higher chlorophyll content and resulting increase in photochemical quenching (qP), apparent electron transport rate, and effective quantum yield of photosystem II (PSII). Both cultivars showed photodamage at 25%FC, but XN889 showed less photodamage in terms of maximal PSII photochemical efficiency. XN889 showed higher qP and non-photochemical quenching than CY17, further demonstrating its superior drought tolerance.     

Spatial and Temporal Variation in Photon Flux Density on Rice (Oryza sativa L.) Leaf Surface

Abstract

Spatial and temporal variations of photosynthetic photon flux density (PFD) measured using small photodiodes (Hamamatsu, model G1118) attached on the leaf surface of rice (Oryza sativa L.) differed with the orientation and inclination even at the same height of the canopy. Under sunny conditions, the fluctuation pattern of PFD was mainly determined by the orientation and inclination of the leaf surface, and not by the daily change of PFD in the open, e. g., high peaks of PFD were observed in the morning on east-oriented leaves, but only low peaks on north-oriented leaves. Under overcast conditions, however, the PFD on the leaf surface depended highly on the PFD in the open irrespective of the leaf orientation and inclination. The present study suggests that the orientation and inclination should be considered as the major factors influencing the PFD regime (spatial variation and daily total PFD) within the rice canopy. To determine the effects of spatial and temporal variations of PFD on crop productivity, the daily net assimilation of each leaf was estimated from the obtained data and the light-photosynthesis curve.     

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...     

不同温光型专用小麦品种花后旗叶生理与籽粒淀粉积累特性

为探索不同温光型专用小麦籽粒淀粉的积累规律及其与植株生长状况的关系,在大田条件下,以2类温光型和3种筋型的小麦品种为材料,研究花后旗叶的生理特性和籽粒淀粉及组分积累情况。结果表明,旗叶中叶绿素含量在灌浆前期(0~21d)维持较高,后期迅速下降,但2个弱筋型品种下降速度缓慢;半冬性品种旗叶中可溶性蛋白含量均大于弱春性品种,而丙二醛(MDA)含量小于弱春性品种,差异均达极显著水平(P0.01)。不同筋型籽粒淀粉组分和总淀粉的积累动态以弱筋型品种最具优势,其直链淀粉含量均在花后14d进入快速增长期,支链淀粉和总淀粉含量在花后28d仍在持续增加,最终以半冬性弱筋品种的籽粒产量和淀粉产量最高(P0.01)。灌浆前期(0~21d),叶绿素、可溶性蛋白含量与直链淀粉、支链淀粉和总淀粉含量呈显著和极显著的相关性(P0.01,P0.05);灌浆后期(21~28d),与直链淀粉的相关性不显著,而叶绿素含量与支链淀粉含量的相关性增大。由此可见,旗叶维持较高的叶绿素、可溶性蛋白含量,较低的丙二醛含量在灌浆前期有利于直链淀粉的积累,后期有利于支链淀粉的积累。     

“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。     

Response of Leaf Photosynthesis to Vapor Pressure Difference in Rice (Oryza sativa L) Varieties in Relation to Stomatal and Leaf Internal Conductance

Abstract

In the afternoon when air humidity decreases, leaf photosynthetic rate (P_n) often declines in rice grown under irrigated conditions. To clarify the genotypic difference of P_n in response to humidity, we measured P_n and stomatal conductance (g_s) for nine rice varieties with diverse genetic backgrounds, at various vapor pressure differences (VPD) and developmental stages. P_n and g_s of all the varieties decreased with VPD increase from 1.0 to 2.3 kPa of VPD. The variety with high g_s at low VPD exhibited a greater decline of g_s with VPD increase than the variety with low g_s, but cv. Takanari showed the highest g_s under altered VPD conditions. Significant logarithmic relations were found between the decreased P_n and g_s at the respective developmental stages, suggesting that g_s is the dominant factor determining P_n and its response to VPD change. To explicate the effect of decreased g_s on P_n, we analyzed the relations by using the model that accurately estimated the genotypic difference in P_n at a low VPD with g_s and leaf nitrogen content per unit leaf area in the previous study. The model assuming that leaf internal conductance (g_w) remains unchanged well explained the decreased P_n at high VPDs by g_s change alone. The analysis also suggested the constancy of g_w and carboxylation capacity at high VPD. It is concluded that the genotypic difference in the decrease of P_n at a high VPD is brought mainly by that in decreased g_s, and the varieties with a high g_s always exhibit a high P_n owing to their relatively high g_s at either high or low VPD environments.     

不同冬小麦品种旗叶叶绿素荧光特性及其对干旱胁迫的响应

为明确不同抗旱性冬小麦品种灌浆期旗叶的叶绿素荧光特性及其与籽粒产量的关系,在防雨棚控水条件下,测定和分析了干旱胁迫后强抗旱性品种晋麦47、弱抗旱性品种偃展4110和中等抗旱性品种矮抗58旗叶荧光参数、籽粒产量和水分利用效率的变化.结果表明,随着先量子密度(PFD)的增加,冬小麦旗叶实际荧光(F)、表观电子传递速率(ETR)和非光化学淬灭系数(NPQ)值逐渐升高,PFD超过189 μmol·m-2·s-2后,F值基本稳定,而光下最大荧光(Fm)值持续降低,导致其他叶绿素荧光参数改变,PFD在189～1 360μtmol·m-2·s-2时Fm变化幅度较大.干旱胁迫降低了小麦旗叶F、Fm、实际量子产量(Yie1d)、ETR、NPQ值.不同品种荧光参数值受干旱胁迫的影响程度不同,偃展4110受影响最大,晋麦47较为稳定.干旱胁迫条件下3个品种的产量差异显著,表现为晋麦47＞矮抗58＞偃展4110.灌浆中期旗叶主要叶绿素荧光参数与产量间具有显著的相关性,可以作为高产品种筛选的生理指标;而干旱处理与适水处理的F比值、Fm比值则与两种水分条件下的产量比值呈显著正相关,可以作为品种抗旱性鉴定的生理指标.