Regulation of reactive oxygen species and phytohormones in osmotic stress tolerance during seed germination in indica rice.

Climate change due to global warming is now affecting agricultural production worldwide. In rice, one of the most important crops, water limitation due to irregular rainfall in rainfed lowlands during crop growth limits yield. Dry direct-sowing has been proposed as a water-efficient approach to cope with water stress during rice growth, but poor seedling establishment due to drought during germination and emergence is a problem. Here, we germinated indica rice cultivars Rc348 (drought tolerant) and Rc10 (drought sensitive) under osmotic stress induced by PEG to elucidate mechanisms of germination under drought. Rc348 had higher germination rate and germination index under severe osmotic stress of -1.5 MPa, above those of Rc10. Rc348 showed up-regulated GA biosynthesis, down-regulated ABA catabolism, and up-regulated α-amylase gene expression in imbibed seeds under PEG treatment compared to that of Rc10. During germination, reactive oxygen species (ROS) play important roles in antagonism between gibberellic acid (GA) and abscisic acid (ABA). Embryo of Rc348 treated with PEG had significantly greater expression of NADPH oxidase genes and higher endogenous ROS levels, together with significantly increased endogenous GA1, GA4 and ABA contents compared to that of Rc10. In aleurone layers treated with exogenous GA, expression of α-amylase genes was higher in Rc348 than in Rc10, and expression of NADPH oxidase genes was enhanced with significantly higher ROS content in Rc348, suggesting higher sensitivity of GA to ROS production and starch degradation in aleurone cells of Rc348. These results suggest that the osmotic stress tolerance of Rc348 is due to enhancement of ROS production, GA biosynthesis, and GA sensitivity, resulting in a higher germination rate under osmotic stress.

Evaluation of the validity of large-scale serial sectioning TEM for three-dimensional reconstruction of rice mesophyll cells and chloroplasts.

Serial sectioning transmission electron microscopy (ssTEM) is a classical method of 3D reconstruction using serial sections obtained with an ultramicrotome. However, producing a long ribbon with homogeneity is difficult. Here, ultramicrotome movement was suspended after producing a ribbon of 15-30 serial sections (cutting intervals, 100 nm), and then, the ribbon was mounted on an individual one-slot grid. However, as this ssTEM method may include influencing factors such as incorrect intervals of section thickness and distortion of sections, which is produced by cutting sections using a diamond knife and beam interaction under TEM observation, qualitative and quantitative data on rice mesophyll cells and chloroplasts were compared with those obtained from a focused ion beam scanning electron microscopy (FIB-SEM) (cutting intervals, 50 nm). No structural distortion in 3D models was observed. In addition, no significant differences in the volume and surface area were observed between the two methods. The surface to volume ratio was significantly affected by the increase in section thickness, but not the difference of methodologies. Our method was useful for observing large volumes of plant cells and organelles, leading to the identification of various sizes and types of chloroplasts. The formation of a chloroplast pocket, which is a structure surrounding other intracellular compartments, was confirmed in rice leaves grown under moderate growth conditions using the ssTEM method. As only four out of 90 chloroplasts formed pocket structures, the formation was considered to be rare under the applied moderate growth conditions.

Three-dimensional ultrastructural change of chloroplasts in rice mesophyll cells responding to salt stress.

BACKGROUND AND AIMS: Excess salinity inhibits the metabolism of various systems and induces structural changes, especially in chloroplasts. Although the chloroplast body seems to swell under salinity stress as observed by conventional transmission electron microscopy, previous studies are limited to 2-D data and lack quantitative comparisons because specimens need to be sliced into ultrathin sections. This study shows three-dimensionally the structural changes in a whole mesophyll cell responding to salinity stress by serial sectioning with a focused ion beam scanning electron microscope (FIB-SEM) and compares the differences in chloroplast structures based on reconstructed models possessing accurate numerical voxel values. METHODS: Leaf blades of rice plants treated with 100 mm NaCl or without (control) for 4 d were fixed chemically and embedded in resin. The specimen blocks were sectioned and observed using the FIB-SEM, and then the sliced image stacks were reconstructed into 3-D models by image processing software. KEY RESULTS: On the transverse sections of rice mesophyll cells, the chloroplasts in the control leaves appeared to be elongated meniscus lens shaped, while those in the salt-treated leaves appear to be expanded oval shaped. The 3-D models based on serial sectioning images showed that the chloroplasts in the control cells spread like sheets fitted to the shape of the cell wall and in close contact with the adjacent chloroplasts. In contrast, those in the salt-stressed cells curled up into a ball and fitted to cell protuberances without being in close contact with adjacent chloroplasts. Although the shapes of chloroplasts were clearly different between the two treatments, their volumes did not differ. CONCLUSIONS: The 3-D reconstructed models of whole rice mesophyll cells indicated that chloroplasts under salt stress conditions were not swollen but became spherical without increasing their volume. This is in contrast to findings of previous studies based on 2-D images.

Influence of Diabetes Mellitus on Surgical Outcomes in Patients with Cervical Myelopathy: A Prospective, Multicenter Study.

STUDY DESIGN: Multicenter, prospective study. PURPOSE: To investigate the effects of diabetes mellitus (DM) on surgical outcomes in patients with cervical myelopathy. OVERVIEW OF LITERATURE: To date, few studies have investigated the influence of postoperative blood glucose or glycated hemoglobin (HbA1c) levels on surgical outcomes. METHODS: The participants were patients who underwent surgery for the treatment of cervical spondylotic myelopathy and ossification of the posterior longitudinal ligament. The 61 cases were evaluated preoperatively and 1 year postoperatively using the Japanese Orthopaedic Association (JOA) scores and the JOA Cervical Myelopathy Evaluation Questionnaire (JOACMEQ). The study variables included fasting blood glucose and HbA1c levels measured preoperatively and at 1 week, 4 weeks, and 1 year postoperatively; the F-wave conduction velocity, latency, rate of occurrence, and M-wave latency in the ulnar and tibial nerves were measured preoperatively and at 1 year postoperatively. The patients were divided into a group without diabetes (N group, 42 patients) and a group with diabetes (DM group, 19 patients). We then assessed the associations between the surgical outcomes and each of the study variables. RESULTS: JOA scores significantly improved in both groups; however, no significant between-group differences were found. There was no significant improvement in the JOACMEQ scores, which assessed cervical function, upper and lower limb function, and bladder function in both groups. We then subdivided the DM group into those with a good control of HbA1c after 1 year (DMG group, 12 patients) and those with HbA1c deterioration after 1 year (DMB group, seven patients), prior to comparing the surgical outcomes. The JOACMEQ scores for upper and lower limb function significantly improved in the DMG group (p<0.01). Compared with the DMB group, there were no significant increases in upper or lower limb function scores in the DMG group. CONCLUSIONS: Poor glycemic control might prevent postoperative functional recovery of the spinal cord.

Three-dimensional ultrastructure of chloroplast pockets formed under salinity stress.

We investigated the invagination structure of a chloroplast that surrounds organelles such as mitochondria and peroxisomes within a thin layer of chloroplast stroma, which is called a chloroplast pocket. In this study, chloroplast pockets were observed in rice plants subjected to salinity stress but not under moderate growth condition. They included cytosol, transparent structure, lipid bodies, mitochondria, and peroxisomes. We constructed the three-dimensional architecture of chloroplast pockets by using serial images obtained by transmission electron microscopy and focused ion beam-scanning electron microscopy. Three types of chloroplast pockets were observed by transmission electron microscopy: Organelles were completely enclosed in a chloroplast pocket (enclosed type), a chloroplast pocket with a small gap in the middle part (gap type), and a chloroplast pocket with one side open (open type). Of the 70 pockets observed by serial imaging, 35 were enclosed type, and 21 and 14 were gap and open types, respectively. Mitochondria and peroxisomes were often in contact with the chloroplast pockets. Focused ion beam-scanning electron microscopy revealed chloroplasts with a sheet structure partially surrounding peroxisomes. This fact suggests that chloroplasts might construct large sheet structures that would be related to the formation of chloroplast pockets.

Three-dimensional intracellular structure of a whole rice mesophyll cell observed with FIB-SEM.

Background and Aims: Ultrathin sections of rice leaf blades observed two-dimensionally using a transmission electron microscope (TEM) show that the chlorenchyma is composed of lobed mesophyll cells, with intricate cell boundaries, and lined with chloroplasts. The lobed cell shape and chloroplast positioning are believed to enhance the area available for the gas exchange surface for photosynthesis in rice leaves. However, a cell image revealing the three-dimensional (3-D) ultrastructure of rice mesophyll cells has not been visualized. In this study, a whole rice mesophyll cell was observed using a focused ion beam scanning electron microscope (FIB-SEM), which provides many serial sections automatically, rapidly and correctly, thereby enabling 3-D cell structure reconstruction. Methods: Rice leaf blades were fixed chemically using the method for conventional TEM observation, embedded in resin and subsequently set in the FIB-SEM chamber. Specimen blocks were sectioned transversely using the FIB, and block-face images were captured using the SEM. The sectioning and imaging were repeated overnight for 200-500 slices (each 50 nm thick). The resultant large-volume image stacks ( x = 25 µm, y = 25 µm, z = 10-25 µm) contained one or two whole mesophyll cells. The 3-D models of whole mesophyll cells were reconstructed using image processing software. Key Results: The reconstructed cell models were discoid shaped with several lobes around the cell periphery. The cell shape increased the surface area, and the ratio of surface area to volume was twice that of a cylinder having the same volume. The chloroplasts occupied half the cell volume and spread as sheets along the cell lobes, covering most of the inner cell surface, with adjacent chloroplasts in close contact with each other. Conclusions: Cellular and sub-cellular ultrastructures of a whole mesophyll cell in a rice leaf blade are demonstrated three-dimensionally using a FIB-SEM. The 3-D models and numerical information support the hypothesis that rice mesophyll cells enhance their CO 2 absorption with increased cell surface and sheet-shaped chloroplasts.

Mixed cropping has the potential to enhance flood tolerance of drought-adapted grain crops.

Recently, the occurrences of extreme flooding and drought, often in the same areas, have increased due to climate change. Wetland plant species are known to oxygenate their rhizospheres by releasing oxygen (O2) from their roots. We tested the hypothesis that wetland species could help upland species under flood conditions; that is, O2 released from the wetland crop roots would ameliorate rhizosphere O2-deficient stress and hence facilitate upland crop root function. Flooding tolerance of upland-adapted staple crops-pearl millet (Pennisetum glaucum) and sorghum (Sorghum bicolor) mix-cropped with rice (Oryza spp.) was investigated in glasshouse and laboratory. We found a phenomenon that strengthens the flood tolerance of upland crops when two species-one wetland and one drought tolerant-were grown using the mixed cropping technique that results in close tangling of their root systems. This technique improved the photosynthetic and transpiration rates of upland crops subjected to flood stress (O2-deficient nutrient culture). Shoot relative growth rates during the flooding period (24 days) tended to be higher under mixed cropping compared with single cropping. Radial oxygen loss from the wetland crop roots might be contributed to the phenomenon observed. Mixed cropping of wet and dryland crops is a new concept that has the potential to overcome flood stress under variable environmental conditions.

Visualization of water transport into soybean nodules by Tof-SIMS cryo system.

This paper examined the route of water supply into soybean nodules through the new visualization technique of time of flight secondary ion mass spectrometry (Tof-SIMS) cryo system, and obtained circumstantial evidence for the water inflow route. The maximum resolution of the Tof-SIMS imaging used by this study was 1.8 µm (defined as the three pixel step length), which allowed us to detect water movement at the cellular level. Deuterium-labeled water was supplied to soybean plants for 4h and the presence of deuterium in soybean nodules was analyzed by the Tof-SIMS cryo system. Deuterium ions were found only in the endodermis tissue surrounding the central cylinder in soybean nodules. Neither xylem vessels nor phloem complex itself did not indicate any deuterium accumulation. Deuterium-ion counts in the endodermis tissue were not changed by girdling treatment, which restricted water movement through the phloem complex. The results strongly indicated that nodule tissues did not receive water directly from the phloem complex, but received water through root cortex apoplastic pathway from the root axis.

First report of Kudoa species in the somatic muscle of the Japanese parrotfish Calotomus japonicus (Scaridae) and a description of Kudoa igami, n. sp. (Myxozoa: Multivalvulida).

Three species of the Kudoid parasite (Myxozoa: Multivalvulida) were observed in the somatic muscle of Japanese parrotfish Calotomus japonicus caught off the coast of western Japan. All three species formed pseudocysts in myofibers and caused subclinical infections. The three Kudoa species were distinguished by spore morphology, as well as their 18S and 28S rDNA sequences. We identified a previously undescribed taxa Kudoa igami n. sp. with spores that were stellate with rounded peripheral edges and five to six polar capsules (prevalence 29.3%). Kudoa igami n. sp. were morphologically most similar to Kudoa neothunni but were distinguishable by a more rounded shape in the apical view. Molecular analyses demonstrated that the K. igami n. sp. is closely related to Kudoa thalassomi; however, the similarity in the 28S rDNA sequence was <96% and the spore morphology was different. We found Kudoa thalassomi in one sample (prevalence 2.4%), which is a new host and geographical record for this species. Kudoa lateolabracis, which causes postmortem myoliquefaction in Chinese sea bass Lateolabrax sp. and olive flounder Paralichthys olivaceus was found in Japanese parrotfish (prevalence 41.5%) for the first time, but did not cause myoliquefaction. We also expanded the host record for the brain-infecting Kudoa yasunagai (prevalence 94.1%). In addition, an unidentified microsporidia was observed in the somatic muscle (prevalence 23.3%).

Early surgical intervention for spinal infection in patients with malignancy requiring chemotherapy: report of two cases and review of the literature.

Advances in chemotherapy for various malignancies have contributed to the increased life expectancy of patients. If such a patient has a concurrent infection, his/her oncologist would hesitate to perform prompt chemotherapy owing to the risk of inducing sepsis. Therefore, the treatment of infection would have priority over initiating chemotherapy for the malignancy. We present a 69-year-old female with malignant lymphoma requiring prompt chemotherapy who also demonstrated spinal infection with Mycobacterium tuberculosis and a 66-year-old male with esophageal cancer who also demonstrated spinal infection with Staphylococcus aureus. Anterior debridement and interbody fusion were performed for both patients. One patient died of malignant lymphoma 4 years after surgery, and the other is still alive and has remained disease-free 4 years after surgery. Saving the life of a patient with malignancy would be difficult without prompt chemotherapy. Conservative treatment for spinal infection requires prolonged antibiotic treatment, and there is no guarantee that the spinal infection would be controlled only with antibiotics. Therefore, early surgical intervention would be an alternative option under such a condition.

Salt-induced chloroplast protrusion is the process of exclusion of ribulose-1,5-bisphosphate carboxylase/oxygenase from chloroplasts into cytoplasm in leaves of rice.

Chloroplast protrusions (CPs) are often observed under environmental stresses, but their role has not been elucidated. The formation of CPs was observed in the leaf of rice plants treated with 75 mm NaCl for 14 d. Some CPs were almost separated from the main chloroplast body. In some CPs, inner membrane structures and crystalline inclusions were included. Similar structures surrounded by double membranes were observed in the cytoplasm and vacuole. Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) was detected in CPs and the similar structures in the cytoplasm and vacuole. These results suggest that CP is one of the pathways of Rubisco exclusion from chloroplasts into the cytoplasm under salinity, and the exclusions could be transported to vacuole for their degradation.

Salinity-induced subcellular accumulation of H(2)O (2) in leaves of rice.

The localization of salt-induced H(2)O(2) accumulation in the leaves of rice was examined using 3,3-diaminobenzidine and CeCl(3) staining at ultrastructure level. When the 3-week-old rice plants were affected by 100 mM NaCl for 14 days, the swelling of thylakoids and the destruction of thylakoid membranes were observed. H(2)O(2) accumulation was also observed in the chloroplast of the leaf treated with NaCl. The electron dense products of 3,3-diaminobenzidine and CeCl(3) were mainly observed especially around the swelling of thylakoids. H(2)O(2) accumulation and any ultrastructural changes were not observed in the chloroplasts under dark condition. Furthermore, treatment with ascorbic acid suppressed both H(2)O(2) accumulation and the changes in chloroplast ultrastructure. These results suggest that light-induced production of excess H(2)O(2) under salinity is responsible for the changes in chloroplast ultrastructure. H(2)O(2) accumulation was also observed in the mitochondria, peroxisomes, plasma membrane, and cell walls under light but not dark, suggesting that these organelles are also the source of H(2)O(2) and the production is light dependent under salinity.

Clinical features and surgical outcomes of cervical spondylotic myelopathy in patients aged 80 years or older: a multi-center retrospective study.

With the aging of the population in developed countries, spine surgeons have recently been more likely to encounter elderly patients in need of treatment. This study investigated whether decompression surgery for cervical spondylotic myelopathy (CSM) in elderly patients aged 80 years or older would likely be a reasonable treatment. We retrospectively reviewed 605 consecutive patients with cervical myelopathy who underwent decompression surgery between 2004 and 2008. Patients with other conditions that could affect functional status or compression factors other than spondylosis were excluded from this study. Of the remaining 189 patients, 161 with CSM whose condition could be evaluated 6 months after surgery were analyzed. The patients were divided into two age groups: 80 years or older (Group A, 37 patients) and younger than 80 years of age (Group B, 124 patients). We evaluated the differences in symptom duration, clinical data, involved levels, surgical outcome, comorbidities, and postoperative complications between the two groups. The symptom duration was significantly shorter in Group A. The average JOA scores preoperatively and 6 months postoperatively were significantly lower in Group A; however, there was no significant difference in the recovery ratio. There were no significant differences in the percentages of patients with comorbidities or those with postoperative complications. Elderly patients aged 80 years or older regained approximately 40% of their function postoperatively, and the incidence of postoperative complication was similar to that in younger patients. Since this age group shows a rapid deterioration after onset, prompt decompression surgery is required.

High-resolution nuclear magnetic resonance spectroscopic study of metabolites in the cerebrospinal fluid of patients with cervical myelopathy and lumbar radiculopathy.

There have been few reports describing substances related to oxidative and intermediary metabolism in the cerebrospinal fluid (CSF) in patients with spinal degenerative disorders. This study investigated whether the concentrations of metabolites in the CSF differed between patients with spinal degenerative disorders and controls, and whether the concentrations of these metabolites correlated with the severity of symptoms. CSF samples were obtained from 30 patients with cervical myelopathy (Group M), 30 patients with lumbar radiculopathy (Group R), and 10 volunteers (control). Metabolites in these CSF samples were measured by nuclear magnetic resonance spectroscopy. There were no differences in the concentrations of lactate, alanine, acetate, glutamate, pyruvate, or citrate between Groups M and R, between Group M and the control, or between Group R and the control. In Group M, neither symptom duration nor the Japanese Orthopaedic Association score correlated with the concentration of any metabolite. In Group R, the symptom duration positively correlated with the concentration of lactate, glutamate, and citrate in CSF. The duration of nerve root block showed a negative correlation with the concentrations of acetate in CSF of the patients in Group R. In patients with lumbar radiculopathy, there is a possibility of increased aerobic metabolic activity or decreased gluconeogenic activity in patients with shorter symptom duration, and increased aerobic metabolic activity in patients with severe inflammation around a nerve root.

Case report: osteoid osteoma of the C2 pedicle: surgical technique using a navigation system.

An osteoid osteoma of the cervical spinal pedicle is rare and carries a high surgical risk because of the close anatomic relationship to the spinal cord, nerve root, and vertebral artery. We report the case of a 12-year-old girl with an osteoid osteoma of the C2 pedicle. Computed tomograms showed an oval nidus and marked sclerosis around this lesion at the right C2 pedicle. There also was expansion of the medial and inferior cortical bone of the C2 pedicle. After failure of nonoperative treatment, we planned surgery. Owing to concerns regarding thermal damage to the spinal cord, nerve root, and/or vertebral artery using computed tomography (CT)-guided radiofrequency ablation, we curetted the nidus using a navigation system. Twenty-eight months after surgery, her pain was relieved with no limitation of cervical movement and there has been no evidence of recurrence. Navigation allowed safe curettage of the nidus through a small hole while maintaining spinal stability.

Recent trends in spinal infections: retrospective analysis of patients treated during the past 50 years.

We retrospectively reviewed the medical charts of 210 consecutive patients treated for spinal infection in Tottori University Hospital in Japan between 1956 and 2005. Until 1995, spinal infection was under control in this series; however, during the last decade, it has been on the rise. Male predominance had been gradually progressing, and 69% of the patients were male during the last decade. Patients with spinal infection were aging, and the ratio of immunocompromised hosts dramatically increased to 53%. Until 1995, the percentage of patients with tuberculous spondylitis had been declining; however, the incidence has been on the rise during the last ten years. The organism was detected in 64% of patients treated between 1996 and 2005, of which Staphylococcus aureus was detected in 49%. Moreover, methicillin-resistant S. aureus was detected in 61% of patients with S. aureus. There were no immigrants or cases with human immunodeficiency virus in this series.

Preferential accumulation of betaine uncoupled to choline monooxygenase in young leaves of sugar beet--importance of long-distance translocation of betaine under normal and salt-stressed conditions.

It has been reported that glycinebetaine (betaine) is synthesized in response to abiotic stresses via a two-step oxidation of choline in which choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH) are involved. Here we show that significant amounts of betaine, > 20 micromol/gFW, accumulated in young leaves of Beta vulgaris even under normal growth conditions, whereas levels in old leaves, cotyledons, hypocotyls, and roots were low. Under the same conditions, CMO accumulates exclusively in old leaves and is difficult to be detected in young leaves. By contrast, the levels of BADH were high in all tissues. Exogenously supplied choline was converted into betaine in old leaves, but levels were significantly lower in young leaves under the same conditions. When d(11)-betaine was applied exogenously to old leaves, it was translocated preferentially into young leaves and roots. In response to salt stress, betaine levels increased in all tissues, but most significantly increased in young leaves. The levels of CMO increased in various tissues, but were low in young leaves. A betaine transporter gene was isolated. Its expression was more strongly induced in old leaves than in young leaves. Based on these data, we discussed the role of CMO and betaine transporter under stress and non-stress conditions.

Tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 in the cerebrospinal fluid of patients with cervical myelopathy and lumbar radiculopathy.

There have been few reports describing cytokines in the cerebrospinal fluid (CSF) of patients with spinal degenerative disorders. This study investigated whether interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) could be detected in CSF of patients with cervical myelopathy or lumbar radiculopathy and whether the concentrations of those cytokines correlated with the severity of disease conditions. CSF samples were obtained from 21 patients with cervical myelopathy (Group M) and 19 patients with lumbar radiculopathy (Group R), and six volunteers (control). The concentration of IL-6 was significantly higher in Groups M and R than in the control, possibly demonstrating spinal cord and nerve root damage, respectively. However, TNF-alpha was lower than the detection limit. IL-1beta was detected in only five samples from three patients in Group M and two volunteers in the control. The concentrations of IL-6 did not show any correlation with symptom duration, the scoring system by the Japanese Orthopaedic Association, or the duration of nerve root block. There is a possibility that the concentration of inflammatory cytokines in CSF can indicate certain pathological aspects of cervical myelopathy or lumbar radiculopathy.

Clinical features and surgical outcomes of cervical myelopathy in the elderly.

UNLABELLED: Treatment of cervical myelopathy in elderly patients is controversial. We retrospectively studied 113 patients who had decompression surgery from 1990-2001 to clarify how pre-operative conditions, duration of symptoms, involved levels, surgical outcomes, and complications differ between younger and elderly patients. We also asked whether elderly patients would likely have reasonable outcomes of surgery. The patients were divided into five age groups: Group 1, 36-45 years (12 patients); Group 2, 46-55 years (22 patients); Group 3, 56-65 years (31 patients); Group 4, 66-75 years (32 patients); and Group 5, 76-85 years (16 patients). The duration of symptoms was similar among the five groups. The involved level was more cephalic in the older groups, and the most frequently involved level in patients older than 75 years was C3-C4. Neurologic symptoms were more severe preoperatively and postoperatively in older patients. The recovery ratio also was lower in older patients; however, in Group 5 it was 36.9%, which indicated that patients older than 75 years could regain approximately 40% of their function postoperatively. Decompression surgery can be a reasonable treatment option for cervical myelopathy, even in elderly patients. LEVEL OF EVIDENCE: Therapeutic Study, Level IV (case series). See the Guidelines for Authors for a complete description of levels of evidence.

Bundle sheath chloroplasts of rice are more sensitive to drought stress than mesophyll chloroplasts.

We investigated the effects of drought stress on the ultrastructure of chloroplasts in rice plants. After the seedlings were grown in a glasshouse for 1 month, they were treated for drought stress using two methods. One drought treatment was imposed by reducing the water supply to the plants for 1 month. The other was imposed by withholding water for 2 weeks to examine the withering process of leaves by drought stress. The ultrastructural changes of chloroplasts in bundle sheath cells were more prominent than those in mesophyll cells under both drought stress treatments. Ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) content in bundle sheath chloroplasts reduced more dramatically than in mesophyll chloroplasts by drought stress. Although a slight swelling of thylakoids was sometimes observed in bundle sheath chloroplasts in moderate stress for 1 month, the thylakoids were less affected by drought stress than chloroplast envelope. These results suggest that chloroplasts in bundle sheath cells were more sensitive to drought stress than those in mesophyll cells and the thylakoids were less damaged by drought stress compared with chloroplast envelope.