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1.
Cédric Jacquard Frédérique Nolin Carine Hécart Dace Grauda Isaak Rashal Sandrine Dhondt-Cordelier Rajbir S. Sangwan Pierre Devaux Florence Mazeyrat-Gourbeyre Christophe Clément 《Plant cell reports》2009,28(9):1329-1339
Albinism remains a major problem in cereal improvement programs that rely on doubled haploid (DH) technology, and the factors
controlling the phenomenon are not well understood. Here we report on the positive influence of copper on the production of
DH plants obtained through microspore embryogenesis (ME) in recalcitrant cultivars of barley (Hordeum vulgare L.). The presence of copper sulphate in the anther pre-treatment medium improved green DH plant regeneration from cultivars
known to produce exclusively albino plants using classical procedures. In plastids, the effect of copper was characterized
by a decrease in starch and a parallel increase in internal membranes. The addition of copper sulphate in the ME pre-treatment
medium should enable breeders to exploit the genetic diversity of recalcitrant cultivars through DH technology. We examined
programmed cell death (PCD) during microspore development to determine whether PCD may interfere with the induction of ME
and/or the occurrence of albinism. By examining the fate of nuclei in various anther cell layers, we demonstrated that the
kinetics of PCD in anthers differed between the barley cultivars Igri and Cork that show a low and a high rate of albinism,
respectively. However, no direct correlation between PCD in the anther cell layers and the rate of albinism was observed and
copper had no influence on the PCD kinetic in these cultivars. It was concluded that albinism following ME was not due to
PCD in anthers, but rather to another unknown phenomenon that appears to specifically affect plastids during microspore/pollen
development. 相似文献
2.
Y. Lee K. Chae J. Ha J. S. Lee I. C. Jang S. Jeong M. Kim M. Yoon 《Russian Journal of Plant Physiology》2009,56(5):654-662
To study gene expression patterns and to find genes related with microspore embryogenesis during pepper (Capsicum annuum L.) anther development, mRNA expression patterns were investigated at four developmental stages distinguished according to
the size of flower bud, the color of anthers, and the cytological feature of microspores. Through GeneFishing using 120 random
primers, 81 genes were found to be differentially expressed as anthers develop. We directly sequenced seven of them, which
were either up- or down-regulated at stage 2, since microspores at stage 2 are known to be responsive to the induction signals
for microspore embryogenesis. Nucleotide sequence analysis of the isolated differentially expressed genes (DEGs) and the comparison
of these sequences with the GenBank data indicate that DEG13 is a novel gene, which is highly homologous to a stress-related
gene of potato, POACT88 (≈91%) and to alcohol dehydrogenase gene of Arabidopsis (≈70%), whose expression is also tightly related to stresses. In vitro data also showed that DEG13 was more abundantly expressed
in heat-treated microspores than in untreated microspores. Here, we report developmental stage-specific gene expression patterns
during anther development and a novel stress-related gene, DEG13, which may be involved in microspore embryogenesis in response
to heat treatment. 相似文献
3.
Wuzhong Yin Hongxia Yang Yantong Wang Ping Feng Yao Deng Yang Liu Danyang Chen Yijie Ban Weichi Liu Guanghua He Nan Wang 《Phyton》2022,91(4):727-744
The mitogen-activated protein kinase (MAPK) cascade is important in stress signal transduction and plant development. In the present study, we identified a rice (Oryza sativa L.) mutant with reduced fertility, Oryza sativa
mitogen-activated protein kinase 6 (osmapk6), which harbored a mutated MAPK gene. Scanning and transmission
electron microscopy, quantitative RT-PCR analysis, TUNEL assays, RNA in situ hybridization, longitudinal and
transverse histological sectioning, and map-based cloning were performed to characterize the osmapk6 mutant.
The gene OsMAPK6 was expressed throughout the plant but predominantly in the microspore mother cells, tapetal cells, and microspores in the anther sac. Compared with the wild type, the total number of microspores was
reduced in the osmapk6 mutant. The formation of microspore mother cells was reduced in the osmapk6 anther
sac at an early stage of anther development, which was the primary reason for the decrease in the total number of
microspores. Programmed cell death of some tapetal cells was delayed in osmapk6 anthers and affected exine formation in neighboring microspores. These results suggest that OsMAPK6 plays pivotal roles in microspore
mother cell formation and tapetal cell degradation. 相似文献
4.
Microarray Analysis of Gene Expression Involved in Anther Development in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) 总被引:6,自引:0,他引:6
Wang Z Liang Y Li C Xu Y Lan L Zhao D Chen C Xu Z Xue Y Chong K 《Plant molecular biology》2005,58(5):721-737
In flowering plants, anthers bear male gametophytes whose development is regulated by the elaborate coordination of many genes. In addition, both gibberellic acid (GA3) and jasmonic acid (JA) play important roles in anther development and pollen fertility. To facilitate the analysis of anther development genes and how GA3 and JA regulate anther development, we performed microarray experiments using a 10-K cDNA microarray with probes derived from seedlings, meiotic anthers, mature anthers and GA3- or JA-treated suspension cells of rice. The expression level change of 2155 genes was significantly (by 2-fold or greater) detected in anthers compared with seedlings. Forty-seven genes, representing genes with potential function in cell cycle and cell structure regulation, hormone response, photosynthesis, stress resistance and metabolism, were differentially expressed in meiotic and mature anthers. Moreover, 314 genes responded to either GA3 or JA treatment, and 24 GA3- and 82 JA-responsive genes showed significant changes in expression between meiosis and the mature anther stages. RT-PCR demonstrated that gene y656d05 was not only highly expressed in meiotic anthers but also induced by GA3. Strong RNA signals of y656d05 were detected in pollen mother cells and tapetum in in situ hybridization. Further characterization of these candidate genes can contribute to the understanding of the molecular mechanism of anther development and the involvement of JA and GA3 signals in the control of anther development in rice. 相似文献
5.
Modification of cell development in vitro: The effect of colchicine on anther and isolated microspore culture in Brassica napus 总被引:12,自引:0,他引:12
In an attempt to discover the biological basis of microspore derived embryogenesis, the effect of the antimicrotubule agent colchicine on anther and free microspore embryogenesis was investigated. The microtubule inhibitor colchicine promoted embryogenesis from cultured anthers, both with regard to the number of anthers responding and the number of embryos being produced per anther. A similar promotional response was also observed with cultured microspores. Although the parameters for cultured anthers and free microspores differed, administration of the drug for a short period immediately prior to pollen mitosis I seems to exert the maximum promotional effect. Of the five cultivars of Brassica napus studied, all responded to colchicine treatment. However, the drug did release more embryogenic potential in poor-responding varieties (i.e. Lirawell and Optima) than in the highest responding variety (Topas). Colchicine also resulted in increased embryogenic response in microspores cultured at lower temperatures.These results are considered in terms of models proposed to explain the switch in microspore development from a gametophytic to a sporophytic pathway. The use ofcolchicine as agent to promote embryogenesis in previously recalcitrant species other than Brassica is also discussed. 相似文献
6.
Ethylene has been shown to be involved in triggering pathogenesis-related (PR) gene expression mainly in dicotyledonous species; however, less attention has been devoted identifying and characterizing PR genes in rice (Oryza sativa L.), a monocot and a model of cereal crop genera. Here, we demonstrate that ethylene induces at least three important rice PR genes, the PR10, PR1 basic (PR1b), and PR5 genes in rice (cv. Nipponbare) seedling leaf, upon treatment with the ethylene generator, ethephon (ET), in a light-, time- and dose-dependent manner. Induction of these PR genes was partially blocked by cycloheximide (CHX), a eukaryotic cytoplasmic protein synthesis inhibitor, which indicates an involvement of cytoplasmic de novo protein synthesis in their induction. These results clearly indicate a dynamic role for ethylene in PR genes induction in rice. 相似文献
7.
Abstract Considerations about our anther cultures of cultivated plants. – One of the main activities performed at the Casaccia Nuclear Centre, in the framework of a contract between CNEN and the European Communities, centers on the induction of haploid plants by anther culture and the subsequent chromosome doubling in order to obtain completely homozygous diploid plants. In tobacco, it is now possible to obtain haploid plants from any cultivar; we perform in vitro culture of internodes from which homozygous diploid plants are regenerated, taking advantage of natural phenomenon of endopolyploidy. In order to try to generalize this method of producing haploid plants in other plant species, we are studying the mechanism involved in haploid embryogenesis which occurs in vitro in the microspores. Datura, Nicotiana and Atropa are among the genera in which a direct embryogenesis from the microspore is observed; it is interesting to note that all three genera belong to the family Solanaceae and are very rich in alkaloids. In almost all the other cases of in vitro induction of haploids, microspores produce calli from which plantlets can be differentiated, but this way of plant regeneration is less interesting because only few plantlets are obtained and it is not sure that each haploid comes from a single microspore. We examined the factors which could influence the transformation of microspores into embryoids in tobacco, namely: the developmental stage of microspore, the degeneration of tapaetal cells, the genotype of microspore, the composition of cultural media, the physiological conditions of the plant from which the anthers were taken. From a practical point of view, it would be desirable to have informations on methods giving a maximum number of haploid plants from one embryogenic anther and the greatest number of embryogenic anthers from the cultured anthers. Our recent experiments on anther culture in liquid shaken medium have yielded good results (about 7,000 embryoids from 25 embryogenic anthers). Further, we are conducting several experiments in order to synchronize the development of the microspores in the anthers; to this end, we analyse the effect of cold treatment, ionizing radiation and gravity force. Experiments are being performed with other cultivated species, beside tobacco, in order to solve some problems of plant breeding more easily and quickly through haploidy. With the aim of introducing, in cultivated tomato, some desirable characters from the wild species, Lycopersicum peruvianum, (self-incompatibility, disease resistance, simultaneous flowering), we have obtained the interspecific hybrid through in vitro culture of young embryos. Haploid production from this hybrid could allow to quickly obtain various genetic recombinations from these two species. For this purpose we are carrying out anther cultures as well as single microspore cultures. In rice, strawberry and L. peruvianum, several diploid and tetraploid plantlets were obtained from our anther cultures. Work is in progress to ascertain the mode of their origin. 相似文献
8.
The ATP-binding Cassette Transporter OsABCG15 is Required for Anther Development and Pollen Fertility in Rice 总被引:1,自引:0,他引:1
Plant male reproductive development is a complex biological process, but the underlying mechanism is not well understood. Here, we characterized a rice (Oryza sativa L.) male sterile mutant. Based on map‐based cloning and sequence analysis, we identified a 1,459‐bp deletion in an adenosine triphosphate (ATP)‐binding cassette (ABC) transporter gene, OsABCG15, causing abnormal anthers and male sterility. Therefore, we named this mutant osabcg15. Expression analysis showed that OsABCG15 is expressed specifically in developmental anthers from stage 8 (meiosis II stage) to stage 10 (late microspore stage). Two genes CYP704B2 and WDA1, involved in the biosynthesis of very‐long‐chain fatty acids for the establishment of the anther cuticle and pollen exine, were downregulated in osabcg15 mutant, suggesting that OsABCG15 may play a key function in the processes related to sporopollenin biosynthesis or sporopollenin transfer from tapetal cells to anther locules. Consistently, histological analysis showed that osabcg15 mutants developed obvious abnormality in postmeiotic tapetum degeneration, leading to rapid degredation of young microspores. The results suggest that OsABCG15 plays a critical role in exine formation and pollen development, similar to the homologous gene of AtABCG26 in Arabidopsis. This work is helpful to understand the regulatory network in rice anther development. 相似文献
9.
microRNAs involved in auxin signalling modulate male sterility under high‐temperature stress in cotton (Gossypium hirsutum)
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Yuanhao Ding Yizan Ma Nian Liu Jiao Xu Qin Hu Yaoyao Li Yuanlong Wu Sai Xie Longfu Zhu Ling Min Xianlong Zhang 《The Plant journal : for cell and molecular biology》2017,91(6):977-994
10.
The present study involves in vitro androgenesis of Zea mays L. using anther culture. We tested combinations of single factors and their influence on microspore induction. Embryogenic induction of microspores within anthers in in vitro conditions was the best when combination of cold treatment, TIBA (0.1 mg l–1) in media and colchicine (0.02% during first 3 days of culture) was applied, but colchicine alone can be factor, which can stimulate or initiate embryogenesis in anther culture of maize. 相似文献
11.
A genic male sterile Chinese cabbage, Brassica campestris L. ssp. chinensis Makino, was examined using cytological and cytochemical methods to characterize the process of pollen abortion in this plant.
Thick sections of both fertile and sterile anthers at different developmental stages were stained using Toluidine Blue O,
Periodic Acid-Schiff’s (PAS) reaction and Sudan Black B to detect cytochemical changes that may occur in the distribution
of insoluble polysaccharide and lipid storage bodies. Pollen abortion in sterile anthers occurs at an early stage of microspore
development. During early microspore development, reductions in the number of starch grains in the connective tissue of fertile
anthers coincide with the accumulation of starch grains in cells of the anther wall. In the late microspore stage, a large
vacuole forms in the microspore, and tapetal cells synthesize and accumulate lipid droplets. The cellular organization of
tapetal cells in sterile anthers appears similar to that in fertile anthers, except for the absence of lipid droplets in cells
of sterile anthers and diffusely labeled tapetal polysaccharides, suggesting defects in nutrient storage.
Supported by National Natural Science Foundation of CHINA (30170060) 相似文献
12.
This study concerns anther culture and the production of microspore-derived calluses and plants of the opium poppy (Papaver somniferum L.). It was confirmed that growth regulators were necessary for microspore callus production. Cold treatment (7 d at 7°C) of the buds prior to culture lead to a twofold increase in the frequency of responsive anthers and in the number of calluses per 100 anthers plated. Callus was produced from cultured anthers of several genotypes, covering a wide genetic background. Step by step removal of growth regulators from the culture medium promoted organogenesis and plant regeneration. Most regenerated plants were diploid. The overall process of microspore embryogenesis closely resembled that described in previous reports on somatic callus production and plant regeneration from poppy hypocotyls in vitro. 相似文献
13.
Maize male reproductive development is complex and lengthy, and anther formation and pollen maturation are precisely and spatiotemporally
regulated. Here, we document that callose, somatic, and microspore defect 1 (csmd1), a new male-sterile mutant, has both pre-meiotic somatic and post-meiotic gametophyte and somatic defects. Chromosome behavior
and cell developmental events were monitored by nuclear staining viewed by bright field microscopy; cell dimensions were charted
by Volocity analysis of confocal microscopy images. Aniline blue staining and quantitative assays were performed to record
callose deposition, and expression of three callose synthase genes was measured by qRT-PCR. Despite numerous defects and unlike
other maize male-sterile mutants that show growth arrest coincident with locular defects, csmd1 anther elongation is nearly normal. Pre-meiotically and during prophase I, there is excess callose surrounding the meiocytes.
Post-meiotically csmd1 epidermal cells have impaired elongation but excess longitudinal divisions, and uninucleate microspores cease growth; the
microspore nucleoli degrade followed by cytoplasmic vacuolization and haploid cell collapse. The single vascular bundle within
csmd1 anthers senesces precociously, coordinate with microspore death. Although csmd1 anther locules contain only epidermal and endothecial cells at maturity, locules are oval rather than collapsed, indicating
that these two cell types suffice to maintain an open channel within each locule. Our data indicate that csmd1 encodes a crucial factor important for normal anther development in both somatic and haploid cells, that excess callose deposition
does not cause meiotic arrest, and that developing pollen is not required for continued maize anther growth. 相似文献
14.
The correlation between the phenologic stage of the inflorescence and the microspore development stage was studied. Cytological
examinations of the development of microspores during in vitro anther culture of cork oak (Quercus suber L.), were carried out during the first four weeks of culture. To observe the division occurring in the microspores, anthers
were taken randomly from the cultures after heat shock treatment and were stained with DAPI. Most of the anthers responding
to a heat stress treatment contained 91 % vacuolated microspores, indicating that this developmental stage is responsive to
embryogenesis induction in cork-oak microspores. After the heat shock treatment some cork-oak microspores were induced and
initiated the embryogenic pathway with the occurrence of numerous symmetric mitosis, producing structures with two to ten
or more nuclei. These lead to the formation of high numbers of multicellular cork-oak microspores (pro-embryos). Twenty-forty
days after induction, small white globular and cotyledonal embryos were observed, which further developed root and shoot,
regenerating plantlets. 相似文献
15.
Iwona Żur Ewa Dubas Elżbieta Golemiec Magdalena Szechyńska-Hebda Gabriela Gołębiowska Maria Wędzony 《Plant cell reports》2009,28(8):1279-1287
Isolated microspore cultures of two spring triticale (x Triticosecale Wittm.) cultivars were used to examine the effect of various stress treatments (either high—32°C or low—5°C temperature with
or without nitrogen/carbohydrate starvation) applied to excised anthers on the effectiveness of microspore embryogenesis induction.
To quantify the effects of pretreatment conditions, the activity of antioxidative enzymes (catalase, peroxidase and superoxide
dismutase) together with respiration rate and heat emission were measured. It was observed that heat shock treatment applied
as the only one stress factor increased the activity of antioxidative enzymes which suggests intensive generation of reactive
oxygen species. Such pretreatment effectively triggered microspore reprogramming but drastically decreased microspore viability.
After low temperature treatment, the activity of antioxidative enzymes was similar to the control subjected only with the
stress originated from the transfer to in vitro culture conditions. This pretreatment decreased the number of microspores
entering embryogenesis but sustained cell viability and this effect prevailed in the final estimation of microspore embryogenesis
effectiveness. For both, low- and high-temperature treatments, interaction with starvation stress was beneficial increasing
microspore viability (at 5°C) or efficiency of embryogenesis induction (at 32°C). The latter treatment significantly reduced
cell metabolic activity. Physiological background of these effects seems to be different and some hypothetical explanations
have been discussed. Received data indicate that in triticale, anther preculture conditions could generate oxidative stress
and change the cell metabolic activity which could next be reflected in the cell viability and the efficiency of microspore
embryogenesis. 相似文献
16.
Early signs of disruption of wheat anther development associated with the induction of male sterility by meiotic-stage water deficit 总被引:10,自引:0,他引:10
Water deficit during meiosis in microspore mother cells of wheat (Triticum aestivum L.) induces male sterility, which reduces grain yield. In plants stressed during meiosis and then re-watered, division of
microspore mother cells seems to proceed normally, but subsequent pollen development is arrested. Stress-affected anthers
generally lack starch. We employed light microscopy in conjunction with histochemistry to compare the developmental anatomy
of water-stress-affected and normal anthers. The earliest effects of stress, detectable between meiosis and young microspore
stages, were the degeneration of meiocytes, loss of orientation of the reproductive cells, and abnormal vacuolization of tapetal
cells. Other effects observed during subsequent developmental stages were deposition of starch in the connective tissue where
it is normally not present, hypertrophy of the middle layer or endothecial cells, and deposition of sporopollenin-like substances
in the anther loculus. The resulting pollen grains lacked both starch and intine. These results suggest that abnormal degeneration
of the tapetum in water-stressed anthers coupled with a loss of orientation of the reproductive cells could be part of early
events leading to abortion of microspores.
Received: 19 July 1996 / Revision accepted: 6 November 1996 相似文献
17.
Mahboobeh Ziaei Mostafa Motallebi Mohammad Reza Zamani Nasim Zarin Panjeh Zahra Moghaddassi Jahromi 《Journal of plant biochemistry and biotechnology.》2016,25(4):358-366
Canola (Brassica napus L.), an agro-economically important crop in the world, is sensitive to many fungal pathogens. One strategy to combat fungal diseases is genetic engineering through transferring genes encoding the pathogenesis-related (PR) proteins such as chitinase which cause the chitin degradation of fungal cell wall. Chitinase Chit42 from Trichoderma atroviride (PTCC5220) plays an important role in biocontrol and has high antifungal activity against a wide range of phytopathogenic fungi. This enzyme lacks a chitin binding domain (ChBD) which is involved in binding activity to insoluble chitin. In the present study, we investigated the effect of chitin binding domain fused to Chit42 when compared with native Chit42. These genes were over-expressed under the CaMV35S promoter in B. napus, R line Hyola 308. Transformation of cotyledonary petioles was achieved by pBISM2 and pBIKE1 constructs containing chimeric and native Chit42 genes respectively, via Agrobacterium method. The insertion of transgenes in T0 generation was verified through polymerase chain reaction (PCR) and Southern blot analysis. Antifungal activity of expressed chitinase in transgenic plants was also investigated by bioassays. The transgenic canola expressing chimeric chitinase showed stronger inhibition against phytopathogenic fungi that indicates the role of chitin binding domain. 相似文献
18.
Improved plant regeneration from wheat anther and barley microspore culture using phenylacetic acid (PAA) 总被引:5,自引:0,他引:5
Summary The effect of the auxin phenylacetic acid (PAA) on wheat anther and on barley anther/microspore culture was investigated. With PAA the induction response was not usually significantly different from controls but a significantly higher number of green plants were produced in wheat anther and barley microspore culture. For wheat anther culture 100 mg/L PAA was beneficial. For barley microspore culture the optimum levels were from 1 to 100 mg/L, depending on genotype. In barley anther culture there were no improvements using PAA. In wheat anther culture, 145 green plants/100 anthers were obtained with cultivar VeeryS, while the average response from twelve F1 hybrids in the breeding program was 332 green plants/100 anthers. At least 1000 green plants were obtained using isolated microspores from 100 anthers in barley cv. Igri. With cv. Bruce, regeneration occurred only when 100 mg/L PAA was used. The influence of PAA appears at the embryogenic phase of the culture system. The possible mechanisms by which PAA may improve regeneration are discussed. 相似文献
19.
20.
C. Halldén G. Karlsson C. Lind I. M. Moller W. K. Heneen 《Sexual plant reproduction》1991,4(3):215-225
Summary The development of sporogenous and tapetal cells in the anthers of male-fertile and cytoplasmic male-sterile sugar beet (Beta vulgaris L.) plants was studied using light and transmission electron microscopy. In general, male-sterile anthers showed a much greater variability in developmental pattern than male-fertile anthers. The earliest deviation from normal anther development was observed to occur in sterile anthers at meiotic early prophase: there was a degeneration or irregular proliferation of the tapetal cells. Other early aberrant events were the occurrence of numerous small vesicles in the microspore mother cells (MMC) and a disorganized chromatin condensation. Deviations that occurred in sterile anthers at later developmental stages included: (1) less distinct inner structures in the mitochondria of both MMC and tapetal cells from middle prophase onwards. (2) dilated ER and nuclear membranes at MMC prophase, in some cases associated with the formation of protein bodies. (3) breakdown of cell walls in MMCs and tapetal cells at late meiotic prophase. (4) no massive increase in tapetal ER at the tetrad stage. (5) a general dissolution of membranes, first in the MMC, then in the tapetum. (6) abortion of microspores and the occurrence of a plasmodial tapetum in anthers reaching the microspore stage. (7) no distinct degeneration of tapetal cells after microspore formation. Thus, it seems that the factors that lead to abortive microsporogenesis are structurally expressed at widely different times during anther development. Aberrant patterns are not restricted to the tetrad stage but occur at early prophase. 相似文献