Genetic control of male fertility in Arabidopsis thaliana : structural analysis of postmeiotic developmental mutants

Seven new male-sterile mutants (ms7–ms13) of Arabidopsis thaliana (L.) Heynh. (ecotype columbia) are described that show a postmeiotic defect of microspore development. In ms9 mutants, microspores recently released from the tetrad appear irregular in shape and are often without exines. The earliest evidence of abnormality in ms12 mutants is degeneration of microspores that lack normal exine sculpturing, suggesting that the MS12 product is important in the formation of pollen exine. Teratomes (abnormally enlarged microsporocytes) are also occasionally present and each has a poorly developed exine. In ms7 mutant plants, the tapetal cytoplasm disintegrates at the late vacuolate microspore stage, apparently causing the degeneration of microspores and pollen grains. With ms8 mutants, the exine of the microspores appears similar to that of the wild type. However, intine development appears impaired and pollen grains rupture prior to maturity. In ms11 mutants, the first detectable abnormality appears at the mid to late vacuolate stage. The absence of fluorescence in the microspores and tapetal cells after staining with 4′,6-diamidino-2-phenylindole (DAPI) and the occasional presence of teratomes indicate degradation of DNA. Viable pollen from ms10 mutant plants is dehisced from anthers but appears to have surface abnormalities affecting interaction with the stigma. Pollen only germinates in high-humidity conditions or during in-vitro germination experiments. Mutant plants also have bright-green stems, suggesting that ms10 belongs to the eceriferum (cer) class of mutants. However, ms10 and cer6 are non-allelic. The ms13 mutant has a similar phenotype to ms10, suggesting is also an eceriferum mutation. Each of these seven mutants had a greater number of flowers than congenic male-fertile plants. The non-allelic nature of these mutants and their different developmental end-points indicate that seven different genes important for the later stages of pollen development have been identified.

Development of plastids in pollen and tapetum of rye-grass, Lolium perenne L

Proplastids of both tapetal cells and microsporocytes were present early in anther development. Tapetal proplastids differentiated—probably into elaioplasts—at late microspore stage. The tapetal cytoplasm was completely resorbed by early tricellular pollen stage. Microspore proplastids differentiated into amyloplasts at early bicellular stage, and were present in both vegetative and generative cells. In the generative cell, the amyloplasts were ephemeral and apparently degenerated within autophagic vacuoles. Plastids were absent from sperm cells. Vegetative cell amyloplasts increased in number apparently by fission such that one amyloplast produced one amyloplast and one proplastid per division. Mature pollen grains were estimated to contain between 550 and 820 amyloplasts with only one starch granule per plastid.

Embryology and seed development of Blastocaulon scirpeum and Paepalanthus scleranthus (Eriocaulaceae)

The embryology and seed structure of Blastocaulon scirpeum (Mart.) Giul. and Paepalanthus scleranthus Ruhland were studied in order to contribute to the embryology of Eriocaulaceae and supply data for future taxonomic studies. Both species present: anther with 4-layered wall; conspicuous endothecium with fibrous thickenings; secretory tapetum with uninucleate cells; successive microsporogenesis forming isobilateral microspore tetrads; bicellular pollen grains; orthotropous, bitegmic and tenuinucellate ovule; micropyle formed by the inner integument alone; megagametophyte of the Polygonum type, with a conspicuous antipodal cyst; nuclear and starchy endosperm; reduced, undifferentiated, and bell-shaped embryo; operculate and endotestal seed; seed coat derived from the two ovule integuments; and tanniniferous endotegmen. In addition, Blastocaulon scirpeum shows a bisporangiate anther and a 3-layered ovary wall, while P. scleranthus presents a tetrasporangiate anther that becomes bisporangiate at maturity, and a 2-layered ovary wall. This investigation shows that the bisporangiate condition does not suffice to separate Blastocaulon from Paepalanthus, since it is common to both. It also indicates, based on several embryological aspects, the proximity of Eriocaulaceae and Xyridaceae, which comply mainly with the features presented by the other commelinid families. These results may be used in future cladistic analysis of the family, and contribute to a better understanding of its phylogeny.

Embryology and seed development of Syngonanthus caulescens (Poir.) Ruhland (Eriocaulaceae-Poales)

The embryology and the seed development of Syngonanthus caulescens are presented. This species possesses: a bithecous and tetrasporangiate anther, with a four-layered wall, a conspicuous endothecium of the baseplate type, a secretory tapetum formed by uninucleate cells, successive microsporogenesis resulting in isobilateral microspore tetrads, spiraperturate and binucleate pollen grains, an orthotropous, pendulous, bitegmic and terminucellate ovule, with a micropyle formed only by the inner integument, a megagametophyte of the Polygonum type, with formation of an antipodal cyst, free-nuclear and starchy endosperm, a broad and bell-shaped embryo, operculate and endotestal seeds, a seed coat derived from the inner layers of both integuments, and tanniniferous endotegmen. These embryological aspects are characteristic not only for Syngonanthus, but for the whole family, with few differences between genera. Furthermore, the pollen grain of the spiraperturate type and the cystic arrangement of the antipodals in the megagametophyte are peculiar and very distinctive features of Eriocaulaceae within the other Poales (commelinids). (c) 2006 Elsevier B.V. All rights reserved.

Embryo-Specific Gene Expression in Microspore-Derived Embryos of Brassica napus. An Interaction between Abscisic Acid and Jasmonic Acid1, 2

The induction of napin and oleosin gene expression in Brassica napus microspore-derived embryos (MDEs) was studied to assess the possible interaction between abscisic acid (ABA) and jasmonic acid (JA). Napin and oleosin transcripts were detected sooner following treatment with ABA than JA. Treatment of MDEs with ABA plus JA gave an additive accumulation of both napin and oleosin mRNA, the absolute amount being dependent on the concentration of each hormone. Endogenous ABA levels were reduced by 10-fold after treatment with JA, negating the possibility that the observed additive interaction was due to JA-induced ABA biosynthesis. Also, JA did not significantly increase the uptake of [3H-ABA] from the medium into MDEs. This suggests that the additive interaction was not due to an enhanced carrier-mediated ABA uptake by JA. Finally, when JA was added to MDEs that had been treated with the ABA biosynthesis inhibitor fluridone, napin mRNA did not increase. Based on these results with the MDE system, it is possible that embryos of B. napus use endogenous JA to modulate ABA effects on expression of both napin and oleosin. In addition, JA could play a causal role in the reduction of ABA that occurs during late stages of seed development.

Genotypic variation for cold tolerance during reproductive development in rice: Screening with cold air and cold water

Rice (Oryza sativa L.) plants are susceptible to low temperature during the young microspore stage, which occurs 10-12 days before heading. Low temperature at this time increases spikelet sterility which can cause massive yield loss. Increasing the cold tolerance of cultivars can reduce yield variability in temperate rice-growing environments. Two experiments were conducted in cold air screenings and two were conducted in cold water screenings to examine genotypic variation for cold tolerance, explore flowering traits related to spikelet sterility, and investigate whether the results reflect the level of cold tolerance determined previously in the field. Cold air screenings imposed day/night temperatures of 27 degrees C/13 degrees C, 25 degrees C/15 degrees C and 32 degrees C/25 degrees C following particle initiation until 50% heading, while cold water screenings maintained a relatively constant 19 degrees C. The variation in the commencement of low air temperature treatment did not have an effect on the level of spikelet sterility, indicating that exposure to low temperature during the young microspore stage was more important than the duration of exposure. Spikelet sterility of common cultivars showed a significant correlation between cold air and cold water screenings (r(2) = 0.63, p < 0.01), cold air and field screenings (r(2) = 0.52, p < 0.01) and cold water and field screenings (r(2) = 0.53, p < 0.01), indicating that cold air and cold water can be used for screening genotypes for low temperature tolerance. HSC55, M 103 and Jyoudeki were identified as cold tolerant and Doongara, Sasanishiki and Nipponbare as susceptible cultivars. There was a significant negative relationship between spikelet sterility and both the number of engorged pollen grains per anther and anther area only after imposing cold air and cold water treatment hence, it was concluded that these flowering traits were facultative in nature. In addition, cultivars originating from Australia and California were inefficient at producing filled grain with similar sized anthers containing a similar number of engorged pollen grains as cultivars from other origins. One suggested reason for this poor conversion to filled grain of cultivars from Australia and California may be associated with their small stigma area, particularly when exposed to low temperature conditions. (c) 2006 Elsevier B.V. All rights reserved.

Minimising cold damage during reproductive development among temperate rice genotypes. I. Avoiding low temperature with the use of appropriate sowing time and photoperiod-sensitive varieties

Multiple-sown field trials in 4 consecutive years in the Riverina region of south-eastern Australia provided 24 different combinations of temperature and day length, which enabled the development of crop phenology models. A crop model was developed for 7 cultivars from diverse origins to identify if photoperiod sensitivity is involved in determining phenological development, and if that is advantageous in avoiding low-temperature damage. Cultivars that were mildly photoperiod-sensitive were identified from sowing to flowering and from panicle initiation to flowering. The crop models were run for 47 years of temperature data to quantify the risk of encountering low temperature during the critical young microspore stage for 5 different sowing dates. Cultivars that were mildly photoperiod-sensitive, such as Amaroo, had a reduced likelihood of encountering low temperature for a wider range of sowing dates compared with photoperiod-insensitive cultivars. The benefits of increased photoperiod sensitivity include greater sowing flexibility and reduced water use as growth duration is shortened when sowing is delayed. Determining the optimal sowing date also requires other considerations, e. g. the risk of cold damage at other sensitive stages such as flowering and the response of yield to a delay in flowering under non-limiting conditions. It was concluded that appropriate sowing time and the use of photoperiod-sensitive cultivars can be advantageous in the Riverina region in avoiding low temperature damage during reproductive development.

Minimising cold damage during reproductive development among temperate rice genotypes. II. Genotypic variation and flowering traits related to cold tolerance screening

Low temperature during microspore development increases spikelet sterility and reduces grain yield in rice (Oryza sativa L.). The objectives of this study were to determine genotypic variation in spikelet sterility in the field in response to low-temperature and then to examine the use of physio-morphological traits at flowering to screen for cold tolerance. Multiple-sown field experiments were conducted over 4 consecutive years in the rice-growing region of Australia to increase the likelihood of encountering low-temperature during microspore development. More than 50 cultivars of various origins were evaluated, with 7 cultivars common to all 4 years. The average minimum temperature for 9 days during microspore development was used as a covariate in the analysis to compare cultivars at a similar temperature. The low-temperature conditions in Year 4 identified cold-tolerant cultivars such as Hayayuki and HSC55 and susceptible cultivars such as Sasanishiki and Doongara. After low temperature conditions, spikelet sterility was negatively correlated with the number of engorged pollen grains, anther length, anther area, anther width, and stigma area. The number of engorged pollen grains and anther length were found to be facultative traits as their relationships with spikelet sterility were identified only after cold water exposure and did not exist under non-stressed conditions.