Tapetal Cells Research Articles

The Arabidopsis cytosolic Acyl-CoA-binding proteins play combinatory roles in pollen development.

In Arabidopsis, six acyl-CoA-binding proteins (ACBPs) have been identified and they have been demonstrated to function in plant stress responses and development. Three of these AtACBPs (AtACBP4-AtACBP6) are cytosolic proteins and all are expressed in floral organs as well as in other tissues. The roles of cytosolic AtACBPs in floral development were addressed in this study. To this end, a T-DNA insertional knockout mutant of acbp5 was characterized before use in crosses with the already available acbp4 and acbp6 T-DNA knockout mutants to examine their independent and combinatory functions in floral development. The single-gene knockout mutations did not cause any significant phenotypic changes, while phenotypic deficiencies affecting siliques and pollen were observed in the double mutants (acbp4acbp6 and acbp5acbp6) and the acbp4acbp5acbp6 triple mutant. Vacuole accumulation in the acbp4acbp6, acbp5acbp6 and acbp4acbp5acbp6 pollen was the most severe abnormality occurring in the double and triple mutants. Furthermore, scanning electron microscopy and transmission electron microscopy revealed exine and oil body defects in the acbp4acbp5acbp6 mutant, which also displayed reduced ability in in vitro pollen germination. Transgenic Arabidopsis expressing β-glucuronidase (GUS) driven from the various AtACBP promoters indicated that AtACBP6pro::GUS expression overlapped with AtACBP4pro::GUS expression in pollen grains and with AtACBP5pro::GUS expression in the microspores and tapetal cells. Taken together, these results suggest that the three cytosolic AtACBPs play combinatory roles in acyl-lipid metabolism during pollen development.

Autophagy in plants and algae.

Autophagy in plants and algae.

ABCG26-mediated polyketide trafficking and hydroxycinnamoyl spermidines contribute to pollen wall exine formation in Arabidopsis.

Pollen grains are encased by a multilayered, multifunctional wall. The sporopollenin and pollen coat constituents of the outer pollen wall (exine) are contributed by surrounding sporophytic tapetal cells. Because the biosynthesis and development of the exine occurs in the innermost cell layers of the anther, direct observations of this process are difficult. The objective of this study was to investigate the transport and assembly of exine components from tapetal cells to microspores in the intact anthers of Arabidopsis thaliana. Intrinsically fluorescent components of developing tapetum and microspores were imaged in intact, live anthers using two-photon microscopy. Mutants of ABCG26, which encodes an ATP binding cassette transporter required for exine formation, accumulated large fluorescent vacuoles in tapetal cells, with corresponding loss of fluorescence on microspores. These vacuolar inclusions were not observed in tapetal cells of double mutants of abcg26 and genes encoding the proposed sporopollenin polyketide biosynthetic metabolon (ACYL COENZYME A SYNTHETASE5, POLYKETIDE SYNTHASE A [PKSA], PKSB, and TETRAKETIDE α-PYRONE REDUCTASE1), providing a genetic link between transport by ABCG26 and polyketide biosynthesis. Genetic analysis also showed that hydroxycinnamoyl spermidines, known components of the pollen coat, were exported from tapeta prior to programmed cell death in the absence of polyketides, raising the possibility that they are incorporated into the exine prior to pollen coat deposition. We propose a model where ABCG26-exported polyketides traffic from tapetal cells to form the sporopollenin backbone, in coordination with the trafficking of additional constituents, prior to tapetum programmed cell death.

DYT1 directly regulates the expression of TDF1 for tapetum development and pollen wall formation in Arabidopsis.

The tapetum plays a critical role during the development and maturation of microspores. DYSFUNCTIONAL TAPETUM 1 (DYT1) is essential for early tapetal development. Here, we determined that the promoter region (-550 to -463bp) contains indispensable cis-elements for DYT1 expression. Although DYT1 transcripts can be detected in both meiocytes and tapetal cells, localization of DYT1-GFP demonstrated that DYT1 is strictly located in tapetal cells during microsporogenesis. Chromatin immunoprecipitation (ChIP) analysis revealed that DYT1 directly binds the promoter region of Defective in Tapetal Development and Function 1 (TDF1), a transcription factor essential for tapetum development. When TDF1 driven by the DYT1 promoter is expressed in a dyt1 mutant, the expression of the transcription factors AMS, MS188/MYB80, TEK and MS1 and the pollen wall-related genes are restored. Although the pollen wall is not formed and the microspores are ruptured, DIOC2 staining showed that fatty acids, the precursors of the pollen wall, were synthesized in the transgenic lines. These results indicate that DYT1 regulates the expression of AMS, MS188/MYB80, TEK and MS1 for pollen wall formation, primarily via TDF1.

Secretory Tissues and the Morphogenesis and Histochemistry of Pollinarium in Flowers of Asclepiadeae (Apocynaceae)

Premise of research. Flowers of some Apocynaceae present an unusual synorganization among parts and organs, which has led to the origin of new organs and a type of pollen dispersal and pollination unique to eudicots. The synorganization of androecium and gynoecium allowed the evolution of the pollinarium, a very complex and species-specific structure composed of pollen grains (pollinium) and secretions produced by the style head (translator). The aim of this study was to investigate the structure and activity of the secretory system related to the morphogenesis of pollinaria and the composition of their secretions in flowers of Asclepiadeae.Methodology. Flowers and buds of Asclepias, Gonioanthela, Matelea, and Oxypetalum were fixed, embedded in Paraplast, and sectioned in a rotary microtome for analysis under light microscopy. SEM was performed for additional structural data, and histochemical tests were carried out to reveal the main chemical composition of the secretions involved in pollinaria morphogenesis.Pivotal results. In the four species of Asclepiadeae studied, the style head possesses a palisade secretory epidermis that is responsible for the secretion of the translator, which is composed of a corpusculum and two caudicles. Mucilage, fatty acids, phenolic compounds, and proteins were detected in the corpusculum, whereas only neutral lipids and mucilage were present in the caudicles. In the anthers, tapetal cells are involved in the secretion of lipids that cover the whole pollinium. Just before anthesis, anthers dehisce, and the pollinia from adjacent anthers contact and adhere to the caudicles of the translator, forming the pollinarium.Conclusions. The translator shape is due to the differential activity of the secretory cells, which are spatially and temporarily coordinated, as well as the amount and composition of the secretion and the undulated outline of the secretory surface. The corpusculum and caudicles are composed of different chemical substances, and these features are related to the processes of removal and insertion of pollinia.

Spatial relationships among the cellular tapetum, visual streak and rod density in dogs.

The dog visual system is well suited to dim light conditions due to rod-dominated retina and the reflective tapetum. The topographical distributions of rods and thickness of the tapetum of the dog were quantified in retinal whole mounts stained with thionine, and spatial relationships among the tapetum, rod density and visual streak of high ganglion cell density were elucidated. The relationship between the retina and tapetum was analyzed in parasagittal sections stained with thionine or hematoxylin-eosin. The tapetum was thick in its center, and the thickest part consisted of 9 to 12 tapetal cell layers. Rod density ranged from 200,000 to 540,000/mm2. Maximum rod density was found in the area dorsal to the visual streak, and the density in that area was significantly higher than the rod density in the visual streak and accorded spatially with the thickest part of the tapetum. The horizontal visual streak was found over the horizontal line through the optic disc in the temporal half and extended slightly into the nasal half. The central area of the highest density of ganglion cells was approximately located midway between the nasal and temporal ends of the visual streak. The visual streak was located within the tapetal area, but ventrally to the thick part of the tapetum.

Microsporogenesis, pollen mitosis and pollen tube growth in Gagea villosa (Liliaceae)

In this study, Gagea villosa (Bieb.) Duby was investigated by using light microscopy methods in cytological and cytoembryological respects. Anthers were tetrasporangiate. Anther wall was formed with an epidermis, endothecium, middle layer and tapetum. Tapetum was glandular type and it began to degenerate when microspores released from tetrads. Tapetum cells generally have one or two nuclei. Mitosis seen in tapetum cells was generally normal but micronuclei were found in some of them. Fibrous thickenings were determined in endothecium. Microsporogenesis and pollen mitosis were generally regular. Cytokinesis was successive type. Meiosis in pollen mother cells was asynchronous in one anther locus. Mature pollen grains were 2-celled. Pollen sterility was found to be 24%. Some of the fertile pollen grains, smaller than the normal were seen at the end of the pollen mitosis. Microgametophyte development was examined in vivo and in vitro. Germination ratio of pollen grains in vitro was 4%. Generally swollen pollen tube tips and weak development of some curled pollen tubes were seen. Callose plug formation was seen only in vivo pollen tube growth.

An anther development F-box (ADF) protein regulated by tapetum degeneration retardation (TDR) controls rice anther development.

In this study, we reported that a F-box protein, OsADF, as one of the direct targets of TDR , plays a critical role in rice tapetum cell development and pollen formation. The tapetum, the innermost sporophytic tissue of anther, plays an important supportive role in male reproduction in flowering plants. After meiosis, tapetal cells undergo programmed cell death (PCD) and provide nutrients for pollen development. Previously we showed that tapetum degeneration retardation (TDR), a basic helix-loop-helix transcription factor, can trigger tapetal PCD and control pollen wall development during anther development. However, the comprehensive regulatory network of TDR remains to be investigated. In this study, we cloned and characterized a panicle-specific expression F-box protein, anther development F-box (OsADF). By qRT-PCR and RNA in situ hybridization, we further confirmed that OsADF expressed specially in tapetal cells from stage 9 to stage 12 during anther development. In consistent with this specific expression pattern, the RNAi transgenic lines of OsADF exhibited abnormal tapetal degeneration and aborted microspores development, which eventually grew pollens with reduced fertility. Furthermore, we demonstrated that the TDR, a key regulator in controlling rice anther development, could regulate directly the expression of OsADF by binding to E-box motifs of its promoter. Therefore, this work highlighted the possible regulatory role of TDR, which regulates tapetal cell development and pollen formation via triggering the possible ADF-mediated proteolysis pathway.

Tapetum specific expression of unedited nad3 gene from safflower and targeting the protein into mitochondria induces male sterility in transgenic tobacco plants

The ability of NAD3 (subunit 3 of the NADH ubiquinone oxidoreductase) encoded by unedited nad3 (u-nad3) gene, when targeted into mitochondria, to cause ablation of tapetum was investigated. A gene construct was developed by cloning the coxIV presequence from yeast and the u-nad3 from safflower under the tapetum specific TA29 promoter to target U-NAD3 into mitochondria in the tapetal cell layer. Transgenic tobacco plants were realized with this construct using the Agrobacterium mediated transformation method and confirmed for the presence of the transgene. Among these plants, three were completely sterile and four were semi-sterile. The male sterile plants were morphologically similar to fertile plants, but the anthers were shorter and remained regressed below the stigma surface at anthesis. RT-PCR and western blotting confirmed anther specific expression of u-nad3 in these plants. Stable inheritance of the induced male sterility and its co-segregation with the introduced gene cassette was confirmed in the test cross progeny. This study demonstrates the proof-of-concept that u-nad3 could be used as a candidate to induce transgenic male sterility in plants.

Basic helix-loop-helix transcription factor BcbHLHpol functions as a positive regulator of pollen development in non-heading Chinese cabbage.

Cytoplasmic male sterility (CMS) is a common trait in higher plants, and several transcription factors regulate pollen development. Previously, we obtained a basic helix-loop-helix transcription factor, BcbHLHpol, via suppression subtractive hybridization in non-heading Chinese cabbage. However, the regulatory function of BcbHLHpol during anther and pollen development remains unclear. In this study, BcbHLHpol was cloned, and its tissue-specific expression profile was analyzed. The results of real-time polymerase chain reaction showed that BcbHLHpol was highly expressed in maintainer buds and that the transcripts of BcbHLHpol significantly decreased in the buds of pol CMS. A virus-induced gene silencing vector that targets BcbHLHpol was constructed and transformed into Brassica campestris plants to further explore the function of BcbHLHpol. Male sterility and short stature were observed in BcbHLHpol-silenced plants. The degradation of tapetal cells was inhibited in BcbHLHpol-silenced plants, and nutrients were insufficiently supplied to the microspore. These phenomena resulted in pollen abortion. This result indicates that BcbHLHpol functions as a positive regulator in pollen development. Yeast two-hybrid and bimolecular fluorescence complementation assays revealed that BcbHLHpol interacted with BcSKP1 in the nucleus. This finding suggests that BcbHLHpol and BcSKP1 are positive coordinating regulators of pollen development. Quantitative real-time PCR indicated that BcbHLHpol and BcSKP1 can be induced at low temperatures. Thus, we propose that BcbHLHpol is necessary for meiosis. This study provides insights into the regulatory functions of the BcbHLHpol network during anther development.

Cytochemical study of developing anthers of Amomum villosum Lour.

We analyzed anther development in Amomum villosum Lour. (Zingiberaceae) using the periodic acid-Schiff's technique and Sudan black staining to test for the presence of starch and lipids, respectively. Our analyses showed that microspore mother cells of A. villosum lack typical callose walls, and numerous lipid granules appear in the cells early in development. Some starch granules are present in anther wall cells, but not in tapetal cells. After meiosis, numerous lipid granules remain unchanged in the microspores. During microspore development, some small starch granules first appear in the central cell region, and then the starch granules increase in size. After microspore division, the bicellular pollen grains become filled with starch and lipids, and remain in this state until the pollen grains reach maturity. At anthesis, the anther wall of A. villosum consists of several layers of endothecium cells with an evidently thickened radial wall, and some layers of parenchyma cells containing numerous starch granules.

A flavonoid 3-O-glucoside:2"-O-glucosyltransferase responsible for terminal modification of pollen-specific flavonols in Arabidopsis thaliana.

Flavonol 3-O-diglucosides with a 1→2 inter-glycosidic linkage are representative pollen-specific flavonols that are widely distributed in plants, but their biosynthetic genes and physiological roles are not well understood. Flavonoid analysis of four Arabidopsis floral organs (pistils, stamens, petals and calyxes) and flowers of wild-type and male sterility 1 (ms1) mutants, which are defective in normal development of pollen and tapetum, showed that kaempferol/quercetin 3-O-β-d-glucopyranosyl-(1→2)-β-d-glucopyranosides accumulated in Arabidopsis pollen. Microarray data using wild-type and ms1 mutants, gene expression patterns in various organs, and phylogenetic analysis of UDP-glycosyltransferases (UGTs) suggest that UGT79B6 (At5g54010) is a key modification enzyme for determining pollen-specific flavonol structure. Kaempferol and quercetin 3-O-glucosyl-(1→2)-glucosides were absent from two independent ugt79b6 knockout mutants. Transgenic ugt79b6 mutant lines transformed with the genomic UGT79B6 gene had the same flavonoid profile as wild-type plants. Recombinant UGT79B6 protein converted kaempferol 3-O-glucoside to kaempferol 3-O-glucosyl-(1→2)-glucoside. UGT79B6 recognized 3-O-glucosylated/galactosylated anthocyanins/flavonols but not 3,5- or 3,7-diglycosylated flavonoids, and prefers UDP-glucose, indicating that UGT79B6 encodes flavonoid 3-O-glucoside:2″-O-glucosyltransferase. A UGT79B6-GUS fusion showed that UGT79B6 was localized in tapetum cells and microspores of developing anthers.

Embryology of Biebersteinia (Biebersteiniaceae, Sapindales): characteristics and comparisons with related families.

Biebersteinia, a perennial herb of five species distributed from Greece to Central Asia, was long considered to be placed in, or near Geraniaceae. Recent molecular analyses, however, have shown that the genus is the sole member of the family Biebersteiniaceae in Sapindales (not including Geraniaceae). Here, we report the embryological features of Biebersteinia and provide embryological corroboration for the molecular sapindalean affinities of the genus. We compared its embryology with those of eight other families of Sapindales, as well as with those of the related orders Huerteales, Malvales, and Brassicales. Overall comparisons showed that Biebersteinia fits in Sapindales because of the presence of anther tapetal cells with polyploid nuclear mass and non-fibrous exotegmen. Further, the genus is characterized by three-celled pollen grains, tetrasporic 16-nucleate Penaea-type female gametophyte, unitegmic ovules, pseudoporogamy, and the chalaza shifting its position near the concave side in the post-fertilization stage. A considerable number of autapomorphies, combined with the lack of synapomorphies with other sapindalean families, supports placing Biebersteinia in its own family. Biebersteiniaceae appear to represent an early divergent lineage of Sapindales. Previous descriptions of seed coats, which were considered to have developed from "bitegmic" ovules, were revised.

The cysteine protease CEP1, a key executor involved in tapetal programmed cell death, regulates pollen development in Arabidopsis.

Tapetal programmed cell death (PCD) is a prerequisite for pollen grain development in angiosperms, and cysteine proteases are the most ubiquitous hydrolases involved in plant PCD. We identified a papain-like cysteine protease, CEP1, which is involved in tapetal PCD and pollen development in Arabidopsis thaliana. CEP1 is expressed specifically in the tapetum from stages 5 to 11 of anther development. The CEP1 protein first appears as a proenzyme in precursor protease vesicles and is then transported to the vacuole and transformed into the mature enzyme before rupture of the vacuole. cep1 mutants exhibited aborted tapetal PCD and decreased pollen fertility with abnormal pollen exine. A transcriptomic analysis revealed that 872 genes showed significantly altered expression in the cep1 mutants, and most of them are important for tapetal cell wall organization, tapetal secretory structure formation, and pollen development. CEP1 overexpression caused premature tapetal PCD and pollen infertility. ELISA and quantitative RT-PCR analyses confirmed that the CEP1 expression level showed a strong relationship to the degree of tapetal PCD and pollen fertility. Our results reveal that CEP1 is a crucial executor during tapetal PCD and that proper CEP1 expression is necessary for timely degeneration of tapetal cells and functional pollen formation.

Anther wall and pollen development in Neotropical species-rich Miconia (Melastomataceae)

Miconia is one of the largest exclusively Neotropical genera, inserted in the tribe Miconieae. Although the monophyly of the tribe has recently been recognized, the delimitation of its genera is considered quite arbitrarily defined, mainly due to the great diversity combined with little morphological and genetic characterization of its members. Recent findings have associated this diversity with polyploidy, apomixis and hybridization, mechanisms commonly found in the group. The conventional delimitation of sections in Miconia, which has been largely based on stamen morphology, has proved to be undoubtedly artificial; nevertheless, the potential taxonomic significance of anther wall and pollen ontogenetic characters within the genera Miconia has been little explored. Hence, this study intended to fill that gap in our knowledge, by investigating the anther wall and pollen development in six closely related species of Miconia: M. albicans, M. fallax, M. latecrenata, M. paucidens, M. pepericarpa and M. stenostachya. Routine techniques for both light and electronic microscopy were used to examine anthers and pollen grains in several developmental stages. The species studied here share several character states, such as the remarkable monocotyledonous anther wall development, the persistent endothecium in the mature anthers with no cell wall thickenings, the bilocular dehiscent anther and the psilate pollen exine. This study also reports for the first time bisporangiate anthers, crystals and proteinoplasts in tapetal cells for Melastomataceae, and ultrastructural features of anther and pollen wall for Miconia.

Decreased Pollen Viability and Thicken Pollen Intine in Antisense Silenced Brassica campestris Mutant of BcMF19

Brassica campestris male fertility 19 (BcMF19; GenBank accession number GQ902048.1), a gene that is specially expressed in tapetum cells and microspores during anther development in B. campestris ssp. chinensis, which is learned from the previous in situ hybridization study. In the present study, we constructed antisense-silenced plants of BcMF19 using B. campestris ssp. chinensis to validate this prediction. The morphology of the pistils, long anthers, and short anthers was significantly affected in 35sbcmf19 compared with the control samples. 4′-6-Diamidino-2-phenylindole staining revealed that two generative nuclei and one large vegetative nucleus were not affected in the mutant compared with control. Statistical analysis of Alexander's staining results showed that 96% of the control pollen grains had vitality, whereas only 86% of the mutant pollen grains did. Under scanning electron microscopy, the mutant demonstrated numerous abnormal pollen grains and resembled dried persimmon. The frequency of normal pollen grains was approximately 18%. Under transmission electron microscopy, the pollen intine during the binucleate and mature pollen stages in 35sbcmf19 exhibited abnormal thickening, especially at the germinal furrows, compared with control. In vitro pollen germination test showed that the tips of the mutant pollen tubes transformed into globular alveoli and stopped growing compared with control. On the other hand, in vivo pollen germination test suggested that BcMF19 affected the pollen tube extension in the pistil. These findings indicate that BcMF19 is essential to the pollen development and pollen tube extension of B. campestris ssp. chinensis.

Sporoderm and tapetum ontogeny in Juniperus communis (Cupressaceae). Connective structures between tapetum and microspores

The purpose of this work was to study in detail the successive stages of pollen wall development in Juniperus communis and intimate connection between the microspores and nutritive tapetum with TEM. Another goal was to clear up the mechanisms underlying the developmental processes. The key stages observed are: the appearance of the initial outer glycocalyx layer as a row of spherical units at the early tetrad stage, followed by the emergence of the underlying radially oriented string-like units, resulting in the formation of a reticulate layer. After sporopollenin accumulation, spherical units turn into granules. String-like units start from the microspore plasma membrane, pierce the glycocalyx layer, pass between the primexine granules and through callose jacket and come out to the anther loculus, reaching the tapetal cells. Their substructure and size are similar to viscin threads, but not their function and chemical composition. All the structures, observed in sporoderm development, correspond to subsequent mesophases of a micellar system. The latter develops by self-assembly, under genomic control, regulating increasing concentration of glycoprotein surfactants in the periplasmic space. A thick network of filaments, connecting microspores and tapetum, supplies microspores with necessary nutrients and testifies the opinion that, in the process of development, exine is a living, dynamic system, arranged from plasmodesmata-like units.

OsATG7 is required for autophagy-dependent lipid metabolism in rice postmeiotic anther development

In flowering plants, the tapetum, the innermost layer of the anther, provides both nutrient and lipid components to developing microspores, pollen grains, and the pollen coat. Though the programmed cell death of the tapetum is one of the most critical and sensitive steps for fertility and is affected by various environmental stresses, its regulatory mechanisms remain mostly unknown. Here we show that autophagy is required for the metabolic regulation and nutrient supply in anthers and that autophagic degradation within tapetum cells is essential for postmeiotic anther development in rice. Autophagosome-like structures and several vacuole-enclosed lipid bodies were observed in postmeiotic tapetum cells specifically at the uninucleate stage during pollen development, which were completely abolished in a retrotransposon-insertional OsATG7 (autophagy-related 7)-knockout mutant defective in autophagy, suggesting that autophagy is induced in tapetum cells. Surprisingly, the mutant showed complete sporophytic male sterility, failed to accumulate lipidic and starch components in pollen grains at the flowering stage, showed reduced pollen germination activity, and had limited anther dehiscence. Lipidomic analyses suggested impairment of editing of phosphatidylcholines and lipid desaturation in the mutant during pollen maturation. These results indicate a critical involvement of autophagy in a reproductive developmental process of rice, and shed light on the novel autophagy-mediated regulation of lipid metabolism in eukaryotic cells.

ABORTIVE POLLEN DEVELOPMENT IN RELATION TO EARLY TAPETAL DEGRADATION IN MALE STERILE MANGOSTEEN

Pollen development in the apomictic mangosteen (Garcinia mangostana L.) has not been exhaustively studied. The objective of this study was to establish the developmental sequence and to determine whether the bursting of the tetrads/ microspores at a time corresponding to microspore release was the factor that contributed to male sterility. Flower buds of various sizes were collected and investigated through combined anatomical observations, histochemical studies, and fluorescence microscopy. The developmental stages were categorized as follows; early pollen mother cell (eaPMC), late pollen mother cell (laPMC), tetrads (TD), early microspore (eaM) and late microspore (laM). Recent studies have shown that the tapetum was disintegrated at the laPMC stage resulting in a significant decrease in the number of tetrads. A scattered primexine wall was deposited around the tetrad. A few of the released microspores could be seen but they did not develop into functional pollen grains. In contrast, most tetrads could not be librated and were degraded. The FCR test revealed that the microspores were not viable. After Oil red O staining, lipid stores were detected in all parts of the anther. Accumulations of carbohydrate were rarely detected (PAS reaction) in the anther wall layers, tapetal cells, sporogenous derivatives and loculi. Male sterility is probably caused by the early degeneration of the tapetum combined with a disruption of the formation of tetrads/microspores. Further studies of the sub-cellular level using analysis by transmission electron microscopy (TEM) are still the subject of intensive research.

Reduction of Gibberellin by Low Temperature Disrupts Pollen Development in Rice

Microsporogenesis in rice (Oryza sativa) plants is susceptible to moderate low temperature (LT; approximately 19°C) that disrupts pollen development and causes severe reductions in grain yields. Although considerable research has been invested in the study of cool-temperature injury, a full understanding of the molecular mechanism has not been achieved. Here, we show that endogenous levels of the bioactive gibberellins (GAs) GA4 and GA7, and expression levels of the GA biosynthesis genes GA20ox3 and GA3ox1, decrease in the developing anthers by exposure to LT. By contrast, the levels of precursor GA12 were higher in response to LT. In addition, the expression of the dehydration-responsive element-binding protein DREB2B and SLENDER RICE1 (SLR1)/DELLA was up-regulated in response to LT. Mutants involved in GA biosynthetic and response pathways were hypersensitive to LT stress, including the semidwarf mutants sd1 and d35, the gain-of-function mutant slr1-d, and gibberellin insensitive dwarf1. The reduction in the number of sporogenous cells and the abnormal enlargement of tapetal cells occurred most severely in the GA-insensitive mutant. Application of exogenous GA significantly reversed the male sterility caused by LT, and simultaneous application of exogenous GA with sucrose substantially improved the extent of normal pollen development. Modern rice varieties carrying the sd1 mutation are widely cultivated, and the sd1 mutation is considered one of the greatest achievements of the Green Revolution. The protective strategy achieved by our work may help sustain steady yields of rice under global climate change.