DCET1 Controls Male Sterility Through Callose Regulation, Exine Formation, and Tapetal Programmed Cell Death in Rice

In angiosperms, anther development comprises of various complex and interrelated biological processes, critically needed for pollen viability. The transitory callose layer serves to separate the meiocytes. It helps in primexine formation, while the timely degradation of tapetal cells is essential for the timely callose wall dissolution and pollen wall formation by providing nutrients for pollen growth. In rice, many genes have been reported and functionally characterized that are involved in callose regulation and pollen wall patterning, including timely programmed cell death (PCD) of the tapetum, but the mechanism of pollen development largely remains ambiguous. We identified and functionally characterized a rice mutant dcet1, having a complete male-sterile phenotype caused by defects in anther callose wall, exine patterning, and tapetal PCD. DCET1 belongs to the RNA recognition motif (RRM)-containing family also called as the ribonucleoprotein (RNP) domain or RNA-binding domain (RBD) protein, having single-nucleotide polymorphism (SNP) substitution from G (threonine-192) to A (isoleucine-192) located at the fifth exon of LOC_Os08g02330, was responsible for the male sterile phenotype in mutant dcet1. Our cytological analysis suggested that DCET1 regulates callose biosynthesis and degradation, pollen exine formation by affecting exine wall patterning, including abnormal nexine, collapsed bacula, and irregular tectum, and timely PCD by delaying the tapetal cell degeneration. As a result, the microspore of dcet1 was swollen and abnormally bursted and even collapsed within the anther locule characterizing complete male sterility. GUS and qRT-PCR analysis indicated that DCET1 is specifically expressed in the anther till the developmental stage 9, consistent with the observed phenotype. The characterization of DCET1 in callose regulation, pollen wall patterning, and tapetal cell PCD strengthens our knowledge for knowing the regulatory pathways involved in rice male reproductive development and has future prospects in hybrid rice breeding.

Sequential Deposition and Remodeling of Cell Wall Polymers During Tomato Pollen Development

The cell wall of a mature pollen grain is a highly specialized, multilayered structure. The outer, sporopollenin-based exine provides protection and support to the pollen grain, while the inner intine, composed primarily of cellulose, is important for pollen germination. The formation of the mature pollen grain wall takes place within the anther with contributions of cell wall material from both the developing pollen grain as well as the surrounding cells of the tapetum. The process of wall development is complex; multiple cell wall polymers are deposited, some transiently, in a controlled sequence of events. Tomato (Solanum lycopersicum) is an important agricultural crop, which requires successful fertilization for fruit production as do many other members of the Solanaceae family. Despite the importance of pollen development for tomato, little is known about the detailed pollen gain wall developmental process. Here, we describe the structure of the tomato pollen wall and establish a developmental timeline of its formation. Mature tomato pollen is released from the anther in a dehydrated state and is tricolpate, with three long apertures without overlaying exine from which the pollen tube may emerge. Using histology and immunostaining, we determined the order in which key cell wall polymers were deposited with respect to overall pollen and anther development. Pollen development began in young flower buds when the premeiotic microspore mother cells (MMCs) began losing their cellulose primary cell wall. Following meiosis, the still conjoined microspores progressed to the tetrad stage characterized by a temporary, thick callose wall. Breakdown of the callose wall released the individual early microspores. Exine deposition began with the secretion of the sporopollenin foot layer. At the late microspore stage, exine deposition was completed and the tapetum degenerated. The pollen underwent mitosis to produce bicellular pollen; at which point, intine formation began, continuing through to pollen maturation. The entire cell wall development process was also punctuated by dynamic changes in pectin composition, particularly changes in methyl-esterified and de-methyl-esterified homogalacturonan.

Embryological studies of Magonia pubescens (Dodonaeaeae, Sapindaceae): development of male and female gametophytes in both floral morphs and its phylogenetic implications

Magonia pubescens A.St.-Hil. (Dodonaeaeae, Sapindaceae) is a monoecious species exhibiting two floral morphs, namely staminate flowers, with gynoecium reduced to a pistillode, and morphologically hermaphrodite but functionally pistillate flowers. It presents the basic type of antheral wall development. Microsporogenesis is normal, forming tetrahedral and decussate tetrads. Anatomical differences in anthers between floral morphs become visible at the stage of callose wall degradation and release of tetrads. In staminate flowers, the endothecium develops fibrous thickening, and the two middle layers, the tapetum and the parenchymal septum that separates both locule, are degraded. At dehiscence, permanent calymmate tetrads are released. Magonia is the only genus of the family with this type of pollen unit. In pistillate flowers, the endothecium exhibits fibrous thickening only in three to five cells on the dorsal loculus, and only the inner middle layer collapses. The septum that separates both locules remains unaltered, the stomium is non-functional, mature anthers are indehiscent and show collapsed tetrads. In staminate flowers, the gynoecium is reduced to a tricarpellar pistillode, trilocular, with ovules that degenerate after megasporogenesis. In pistillate flowers, the gynoecium has a tricarpellary ovary, with six to eight ovules per carpel; they are campylotropous, bitegmic, mixed crassinucellate, and exhibit a well-developed obturator. The phylogenetic implications of these embryological characters are discussed in the context of the family.

Ultrastructural transformations in the cytoplasm of differentiating Hyacinthus orientalis L. pollen cells

The sequence of ultrastructural changes in the cytoplasm during the successive stages of pollen grain development in <em>Hyacinthus orientulis</em> pollen cells was studied. The cytoplasmic transformations of the generative cell included the elimination of plastids, increase in the number of mitochondria, assumption of a spindle shape with the aid of microtubules and the characteristic development of the vacuole system with the formation of so-called colored bodies. The cytoplasmic transformations of the generative cell encompassed changes in the plastids, which began to accumulate starch soon after the cell was formed, then released it shortly before anthesis, an increase in the number of mitochondria and an increase in the number of highly active dictyosomes just before anthesis. Changes in the structure of the border region between the differentiating pollen cells were associated mainly with the periodical appearance of a callose wall and the presence of lysosome-like bodies in the cytoplasm of the vegetative cell surrounding the generative cell. They arose soon after the disappearance of the callose wall and disappeared shortly before anthesis.

Development of the pollen in the antarctic flowering plant Colobanthus quitensis (Kunth) Bartl.

<i>Colobanthus quitensis</i> (Kunth) Bartl. produced two types very small bisexual fl owers. In the Antarctic natural conditions chasmogamic and cleistogamic fl owers most often form fi ve stamina with short fi laments. Two microsporangia with a three-layer wall form in the anther. Microspore mother cells, which develop into microspores after meiosis, form inside the microsporangium. Microsporocytes of <i>Colobanthus quitensis</i> are surrounded with a thick callose layer, the special wall. After meiosis, the callose wall is dissolved and microspores are released from the tetrad. The production of proorbicules, orbicules and peritapetal membrane, and the construction of a complex sporoderm with numerous apertural sites were observed. When microspore and pollen protoplasts underwent necrosis, probably as a result of temperature and osmotic stress, sporoderm layers formed around microspores, and the cell tapetum did not disintegrate. However, woody wall layers did not accumulate in endothecium cells.