Showing papers on "Endosperm published in 1992"

The Maize Invertase-Deficient miniature-1 Seed Mutation Is Associated with Aberrant Pedicel and Endosperm Development.

Abstract: Genetic evidence is presented to show that the developmental stability of maternal cells in the pedicel at the base of maize seeds is determined by the genotype of the developing endosperm. An early degeneration and withdrawal of maternal cells from the endosperm of homozygous miniature (mn mn) seed mutants were arrested if mn plants were pollinated by the wild-type Mn pollen. Similarly, the stability of the wild-type, Mn mn, maternal cells was also dependent on whether or not these cells were associated with the normal (Mn) or the mutant (mn) endosperm on the same ear. Biochemical and cellular analyses indicated that developing mn kernels have extremely low (<0.5% of the wild type) to undetectable levels of both soluble and wall-bound invertase activities. Extracts from endosperm with a single copy of the Mn gene showed a significant increase in both forms of invertases, and we suggest it is the causal basis of the wild-type seed phenotype. Collectively, these data provide evidence that invertase-mediated maintenance of a physiological gradient of photosynthate between pedicel and endosperm constitutes the rate-limiting step in structural stability of maternal cells as well as normal development of endosperm and seed.

Starch Hydrolysis by the Ruminal Microflora

Abstract: The effects of grain type and processing on ruminal starch digestion are well documented but poorly understood at the biochemical and molecular levels. Waxy grains have starches high in amylopectin and are more readily digested than nonwaxy grains. However, the composition of the endosperm cell matrix and the extent to which the starch granules are embedded within it also affect starch digestion rates. Continued work is needed to determine the influence of specific cell matrix proteins, protein-starch interactions and cell wall carbohydrates on starch availability. The microbial populations that metabolize starch are diverse, differing in their capacities to hydrolyze starch granules and soluble forms of starch. Surveys show that the amylases are under regulatory control in most of these organisms, but few studies have addressed the types of amylolytic enzymes produced, their regulation and the impact of other plant polymers on their synthesis. Research in these areas, coupled with the development and use of isogeneic or near-isogeneic grain cultivars with biochemically defined endosperm characteristics, will enhance our ability to identify mechanisms to manipulate ruminal starch digestion.

A gibberellin response complex in cereal alpha-amylase gene promoters.

Abstract: The Amy32b gene is a representative member of a closely related family of alpha-amylase genes expressed under hormonal control in aleurone layers of barley grains. Transcription of this gene is induced by gibberellin (GA) and suppressed by abscisic acid. In this study, we functionally defined the promoter elements of the Amy32b gene that govern the developmental and hormonal control of its expression in aleurone. Two functionally distinct yet physically associated elements are essential: a gibberellin response element mediates regulation by GA and abscisic acid, and an Opaque-2 binding sequence (O2S) is thought to interact with a barley homolog of the maize endosperm-specific transcriptional regulator Opaque-2. An additional element CCTTTT, which with the O2S forms part of a canonical "endosperm box," is important in modulating the absolute level of expression of the Amy32b promoter, as is another separate, highly conserved element TATCCATGCAGTG.

Dormancy and Germination of Abscisic Acid-Deficient Tomato Seeds Studies with the sitiens Mutant

Abstract: The role of abscisic acid (ABA) in the dormancy induction of tomato (Lycopersicon esculentum) seeds was studied by comparison of the germination behavior of the ABA-deficient sitiens mutant with that of the isogenic wild-type genotype. Freshly harvested mutant seeds, in contrast to wild-type seeds, always readily germinate and even exhibit viviparous germination in overripe fruits. Crosses between mutant and wild-type and self-pollination of heterozygous plants show that in particular the ABA fraction of embryo and endosperm is decisive for the induction of dormancy. After-ripened wild-type seeds fully germinate in water but are more sensitive toward osmotic inhibition than mutant seeds. Germination of both wild-type and mutant seeds is equally sensitive toward inhibition by exogenous ABA. ABA content of mature wild-type seeds is about 10-fold the level found in mutant seeds. Nevertheless, it is argued that the differences in dormancy between the seeds of both genotypes are not a result of actual ABA levels in the mature seeds or fruits but a result of differences in ABA levels during seed development. It is hypothesized that the high levels of ABA that occur during seed development in wild-type seeds induce an inhibition of cell elongation of the radicle that can still be observed after long periods of dry storage.

Waxy Chlamydomonas reinhardtii: monocellular algal mutants defective in amylose biosynthesis and granule-bound starch synthase activity accumulate a structurally modified amylopectin.

Abstract: Amylose-defective mutants were selected after UV mutagenesis of Chlamydomonas reinhardtii cells. Two recessive nuclear alleles of the ST-2 gene led to the disappearance not only of amylose but also of a fraction of the amylopectin. Granule-bound starch synthase activities were markedly reduced in strains carrying either st-2-1 or st-2-2, as is the case for amylose-deficient (waxy) endosperm mutants of higher plants. The main 76-kDa protein associated with the starch granule was either missing or greatly diminished in both mutants, while st-2-1-carrying strains displayed a novel 56-kDa major protein. Methylation and nuclear magnetic resonance analysis of wild-type algal storage polysaccharide revealed a structure identical to that of higher-plant starch, while amylose-defective mutants retained a modified amylopectin fraction. We thus propose that the waxy gene product conditions not only the synthesis of amylose from endosperm storage tissue in higher-plant amyloplasts but also that of amylose and a fraction of amylopectin in all starch-accumulating plastids. The nature of the ST-2 (waxy) gene product with respect to the granule-bound starch synthase activities is discussed. Images

Effect of anoxia on starch breakdown in rice and wheat seeds

Abstract: The capabilities of rice (Oryza sativa L.) and wheat (Triticum aestivum L.) seeds (caryopses) to degrade starchy reserves present in the endosperm tissue were compared under anaerobic conditions. The results showed that rice, a species highly tolerant to anoxia, can readily break down starch under anaerobiosis concomitant with germination, while wheat does not germinate and fails to degrade starch present in the endosperm. This clearly distinct behavior is likely the consequence of the successful inducible formation of α-amylase (EC 3.2.1.1.) in rice under anoxia, whereas the enzyme is not produced in wheat seeds. We found that rice seeds possess a set of enzymes allowing starch and its degradative products to be utilized under anoxic conditions. Wheat seeds were shown to germinate even under anoxia if fed glucose or sucrose exogenously. The overall results indicate that induction of α-amylase appears to be one of the factors permiting rice seeds to germinate in totally anaerobic environments.

Developmental and organ-specific expression of an ABA- and stress-induced protein in barley.

Abstract: An mRNA species, HVA1, has been shown to be rapidly induced by abscisic acid (ABA) in barley aleurone layers (Hong, Uknes and Ho, Plant Mol Biol 11: 495-506, 1988). In the current work we have investigated the expression of HVA1 in other organs of barley plants. In developing seeds, HVA1 mRNA is not detected in starchy endosperm cells, yet it accumulates in aleurone layers and embryo starting 25 days after anthesis, and its level remains high in these organs in dry seeds. Although the levels of HVA1 mRNA are equivalent in the dry embryos of dormant and nondormant barley seeds, upon imbibition HVA1 mRNA declines much slower in the dormant than in the nondormant embryos. The HVA1 mRNA and protein levels are highly induced by ABA treatment in all organs of 3-day-old seedlings. However, the induction in the leaf of 7-day-old seedlings is less than one tenth the level observed in the leaf of 3-day-old seedlings. In the leaf, HVA1 mRNA and protein are induced mainly at the base. These observations indicate that the expression of HVA1 is under developmental regulation. Besides the HVA1 protein, a smaller protein (p20) of approximately 20 kDa cross-reacting with anti-HVA1 polyclonal antibodies, is induced by ABA in barley seedlings but not in seeds. HVA1 mRNA is induced by drought, NaCl, cold or heat treatment. Similar to ABA treatment, the drought induction of HVA1 occurs in all the tissues of 3-day-old seedlings, but the induction decreases dramatically in the leaf of 7-day-old plants. The significance of organ-specific, developmentally regulated, and stress-induced expression of HVA1 is discussed.

Malate- and pyruvate-dependent Fatty Acid synthesis in leucoplasts from developing castor endosperm.

Abstract: Leucoplasts were isolated from the endosperm of developing castor (Ricinis communis) endosperm using a discontinuous Percoll gradient. The rate of fatty acid synthesis was highest when malate was the precursor, at 155 nanomoles acetyl-CoA equivalents per milligram protein per hour. Pyruvate and acetate also were precursors of fatty acid synthesis, but the rates were approximately 4.5 and 120 times less, respectively, than when malate was the precursor. When acetate was supplied to leucoplasts, exogenous ATP, NADH, and NADPH were required to obtain maximal rates of fatty acid synthesis. In contrast, the incorporation of malate and pyruvate into fatty acids did not require a supply of exogenous reductant. Further, the incorporation of radiolabel into fatty acids by leucoplasts supplied with radiolabeled malate, pyruvate, or acetate was reduced upon coincubation with cold pyruvate or malate. The data suggest that malate and pyruvate may be good in vivo sources of carbon for fatty acid synthesis and that, in these preparations, leucoplast fatty acid synthesis may be limited by activity at or downstream of the acetyl-CoA carboxylase reaction.

Evidence for the presence of two different types of protein bodies in wheat endosperm.

Abstract: Storage proteins of wheat grains (Triticum L. em Thell) are deposited in protein bodies inside vacuoles. However, the subcellular sites and mechanisms of their aggregation into protein bodies are not clear. In the present report, we provide evidence for two different types of protein bodies, low- and high-density types that accumulate concurrently and independently in developing wheat endosperm cells. Gliadins were present in both types of protein bodies, whereas the high molecular weight glutenins were localized mainly in the dense ones. Pulse-chase experiments verified that the dense protein bodies were not formed by a gradual increase in density but, presumably, by a distinct, quick process of storage protein aggregation. Subcellular fractionation and electron microscopy studies revealed that the wheat homolog of immunoglobulin heavy-chain-binding protein, an endoplasmic reticulum-resident protein, was present within the dense protein bodies, implying that these were formed by aggregation of storage proteins within the endoplasmic reticulum. The present results suggest that a large part of wheat storage proteins aggregate into protein bodies within the rough endoplasmic reticulum. Because these protein bodies are too large to enter the Golgi, they are likely to be transported directly to vacuoles. This route may operate in concert with the known Golgi-mediated transport to vacuoles in which the storage proteins apparently condense into protein bodies at a postendoplasmic reticulum location. Our results further suggest that although gliadins are transported by either one of these routes, the high molecular weight glutenins use only the Golgi bypass route.

Partial Purification and Characterization of Lysine-Ketoglutarate Reductase in Normal and Opaque-2 Maize Endosperms

Abstract: Lysine-ketoglutarate reductase catalyzes the first step of lysine catabolism in maize (Zea mays L.) endosperm. The enzyme condenses l-lysine and α-ketoglutarate into saccharopine using NADPH as cofactor. It is endosperm-specific and has a temporal pattern of activity, increasing with the onset of kernel development, reaching a peak 20 to 25 days after pollination, and there-after decreasing as the kernel approaches maturity. The enzyme was extracted from the developing maize endosperm and partially purified by ammonium-sulfate precipitation, anion-exchange chromatography on DEAE-cellulose, and affinity chromatography on Blue-Sepharose CL-6B. The preparation obtained from affinity chromatography was enriched 275-fold and had a specific activity of 411 nanomoles per minute per milligram protein. The native and denaturated enzyme is a 140 kilodalton protein as determined by polyacrylamide gel electrophoresis. The enzyme showed specificity for its substrates and was not inhibited by either aminoethyl-cysteine or glutamate. Steady-state product-inhibition studies revealed that saccharopine was a noncompetitive inhibitor with respect to α-ketoglutarate and a competitive inhibitor with respect to lysine. This is suggestive of a rapid equilibrium-ordered binding mechanism with a binding order of lysine, α-ketoglutarate, NADPH. The enzyme activity was investigated in two maize inbred lines with homozygous normal and opaque-2 endosperms. The pattern of lysine-ketoglutarate reductase activity is coordinated with the rate of zein accumulation during endosperm development. A coordinated regulation of enzyme activity and zein accumulation was observed in the opaque-2 endosperm as the activity and zein levels were two to three times lower than in the normal endosperm. Enzyme extracted from L1038 normal and opaque-2 20 days after pollination was partially purified by DEAE-cellulose chromatography. Both genotypes showed a similar elution pattern with a single activity peak eluted at approximately 0.2 molar KCL. The molecular weight and physical properties of the normal and opaque-2 enzymes were essentially the same. We suggest that the Opaque-2 gene, which is a transactivator of the 22 kilodalton zein genes, may be involved in the regulation of the lysine-ketoglutarate reductase gene in maize endosperm. In addition, the decreased reductase activity caused by the opaque-2 mutation may explain, at least in part, the elevated concentration of lysine found in the opaque-2 endosperm.

Metabolic regulation of rice alpha-amylase and sucrose synthase genes in planta.

Abstract: Summary Isolated rice embryos were used to investigate the regulatory effects of endosperm extracts and pure sugars on the expression of α-amylase gene RAmy3D and a sucrose synthase gene homologous to the maize isozyme Ss2 The high-level expression of RAmy3D in the scutella of isolated embryos could be inhibited by a variety of sugars as well as endosperm extracts from germinated rice grains Glucose, at a concentration of 250 mM, was most effective in repressing RAmy3D mRNA accumulation Furthermore, this repression was reversible Interestingly, RAmy3D repression was always accompanied by the induction of sucrose synthase gene expression These results support a model in which the expression of α-amylase and sucrose synthase genes in the rice scutellum are counter-regulated by the influx of sugars from the endosperm

Endosperm differentiation in barley wild-type andsexmutants

Abstract: The cereal endosperm is a storage organ consisting of the central starchy endosperm surrounded by the aleurone layer. In barley, endosperm development is subdivisible into four main stages, i.e. the syncytial (I), the cellularization (II), the differentiation (III) and the maturation stage (IV). During stage I, a multinucleate syncytium is formed, which in stage II develops into the undifferentiated cellular endosperm. During stage III the cells of the endosperm differentiate into two types of aleurone cells (peripheral and modified) and three different starchy endosperm cell types (irregular, prismatic and subaleurone). To elucidate the ontogenetic relationship between the endosperm tissues, the phenotypes of sex (shrunken endosperm mutants expressing xenia) mutant endosperms were studied. These mutants can be classified into two groups, i.e. those in which development is arrested at one of the four wild-type stages described above, and those with abnormal development with new organizational patterns in the endosperm or with novel cell types. Based on these studies, it is suggested that the two endosperm halves represent cell lines derived from the two daughter nuclei of the primary endosperm nucleus, and that the prismatic starchy endosperm cells arise from a peripheral endosperm meristematic activity during stage III. Finally, a model for the main molecular events underlying the morphogenetic processes is discussed.

Increases in binding protein (BiP) accompany changes in protein body morphology in three high-lysine mutants of maize

Abstract: A maize 75 kDa protein recently has been identified as a plant homolog of the mammalian binding protein (BiP). To better understand the function of BiP in protein body formation in maize endosperm, immunomicroscopy studies were conducted on three maize endosperm mutants, floury-2, Mucronate, and Defective endosperm-B 30, in which the level of BiP is highly elevated. Our results showed that protein body morphology in all three mutants was altered. In addition, BiP was localized in both the ER and peripheral regions of the abnormal protein bodies. The degree to which protein body morphology differed from normal was positively correlated with increased amounts of BiP. In addition, the accumulation of BiP in abnormal protein bodies increased with protein body maturation. In the three endosperm mutants, the arrangement of zeins within protein bodies had been perturbed, yet none of the specific zein subclasses exhibited the staining pattern found for BiP. The association of BiP with abnormal packaging of proteins in protein bodies may reflect a biological function to mediate protein folding and assembly in maize endosperm.

Purification and immunocytochemical localization of kafirins in Sorghum bicolor (L. Moench) endosperm

Abstract: Kafirins are the storage proteins of sorghum and are found in protein bodies in the seed endosperm. They have been classified as α-, β-, and γ-kafirins according to differences in molecular weight, solubility, and structure. The kafirins were purified, amino acid composition was determined, and immunolocalization methods were used to determine the organization of the protein bodies and distribution of kafirins throughout the endosperm. All three groups of kafirins were low in lysine. β-Kafirins and γ-kafirins were relatively high in cysteine, and β-kafirins were relatively high in methionine. Transmission electron microscopy showed that protein bodies in the peripheral endosperm were spheroid with concentric rings and few darkly stained inclusions. In contrast, protein bodies of the central endosperm were irregularly shaped with a higher proportion of darkly stained material. The light staining regions of the protein bodies are composed primarily of α-kafirins with minor portions of β- and γ-kafirins. The dark staining regions, however, are composed primarily of β- and γ-kafirins. Immunoelectron microscopy showed that protein bodies in the peripheral endosperm contain predominantly a-kafirin with minor amounts of β- and γ-kafirin. Central endosperm protein bodies are also predominantly α-kafirin, but have a higher proportion of β-kafirin and γ-kafirin than the peripheral endosperm protein bodies.

Appearance of Endoproteolytic Enzymes during the Germination of Barley

Abstract: Barley endoproteolytic enzymes are important to germination because they hydrolyze endosperm storage proteins to provide precursors for new protein synthesis. We recently developed an electrophoretic method utilizing gel-incorporated protein substrates to study the endoproteinases of 4-d-germinated barley (Hordeum vulgare L. cv Morex) grain. This work extends those findings to determine the temporal pattern of the appearance of the endoproteinases during germination, the sensitivities of the proteinases to class-specific proteinase inhibitors, and where, in germinating caryopses, the proteinases reside. Six endoproteinase activity bands (representing a minimum of seven enzymes) were present in 5-d-germinated barley grain extracts subjected to electrophoresis in nondenaturing gels at pH 8.8. The activities of two of the enzyme bands (“neutral” proteinases) increased as the pH was increased from 3.8 to 6.5. The activities of the remaining four (“acidic”) bands diminished abruptly as the pH increased above 4.7. Two proteinase bands hydrolyzed gelatin but not edestin, four of the proteinases hydrolyzed both gelatin and edestin at nearly the same rates, and one enzyme degraded only edestin. One neutral endoproteinase was sensitive to diisopropyl fluorophosphate inhibition, and the other was not inhibited by any of inhibitors tested. Four of acidic enzymes were cysteine proteinases [inhibited by trans-epoxysuccinyl-l-leucylamido(4-guanidino)butane and N-ethylmaleimide]; the other was an aspartic acid endoproteinase (sensitive to pepstatin). Only the aspartic proteinase was detected in either ungerminated or steeped barley grain. During the germination (malting) process, the aspartic endoproteinase activity decreased until the second day of germination and then increased until germination day 5. The first endoproteinase(s) induced during germination was a neutral enzyme that showed activity on the 1st day of the germination phase after steeping. Most of the endoproteinases became active on the 2nd or 3rd germination day, but one cysteine proteinase was not detected until the 5th day. Acid cysteine proteinases were present in the aleurone, scutellum, and endosperm tissues but not in shoots and roots. The aleurone layer and endosperm contained almost exclusively band B1 neutral proteinases, whereas the scutellum, shoots, and roots contained both B1 and B2 bands. This work shows that germinating barley contains a complex set of proteinases whose expression is temporally and spatially controlled. But, at the same time, it also shows that this electrophoretic method for separating and studying individual enzymes of this complex will allow us to more readily characterize and purify them.

Properties of Barley Seed Chitinases and Release of Embryo-Associated Isoforms during Early Stages of Imbibition

Abstract: Barley (Hordeum vulgare L.) seeds contain at least five proteins with chitinase (CH) activity. Two of these (CH1 and CH2) are found primarily in the aleurone and endosperm tissues, and the other three (CH3, CH4, and CH5) are enriched in the embryo. From the bran fraction, three of these CHs (CH1, CH2, and CH3) were purified to apparent homogeneity. These three CHs have apparent molecular masses of 27, 34, and 35 kilodaltons and isoelectric points of 9.3, 9.2, and 8.7, respectively. CH2 and CH3 have amino terminal sequences resembling a portion of the chitin-binding domain of lectins and other plant defense proteins. CH1 lacks this domain. All three CHs exhibit antifungal activity and inhibit the mycelial growth of some species of trichoderma and Fusarium in vitro. During the early period of imbibition by seeds, two of the embryo-associated CHs are selectively released into the surrounding aqueous medium.

Phosphoenolpyruvate carboxylase activity and concentration in the endosperm of developing and germinating castor oil seeds.

Abstract: Monospecific polyclonal antibodies against maize leaf phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) were utilized to examine the subunit composition and developmental profile of endosperm PEPC in developing and germinating castor oil seeds (Ricinus communis L. cv Baker 296). PEPC from developing endosperm consists of a single type of 100-kilodalton subunit, whereas the enzyme from 2- to 5-day germinated endosperm appears to contain equal proportions of immunologically related 103- and 108-kilodalton subunits. The maximal activity of PEPC in developing endosperms (2.67 micromoles oxaloacetate produced per minute per gram fresh weight) is approximately 20-fold and threefold greater than that of fully mature (dry seed) and germinating endosperms, respectively. The most significant increase in the activity and concentration of endosperm PEPC occurs during the middle cotyledon to full cotyledon stage of seed development; this period coincides with the most active phase of storage oil accumulation by ripening castor oil seeds. The data are compatible with the recent proposal (RG Smith, DA Gauthier, DT Dennis, DH Turpin [1992] Plant Physiol 1233-1238) that PEPC plays a fundamental role in vivo in the cytosolic production of an important substrate (malate) for fatty acid biosynthesis by developing castor oil seed leucoplasts. Immediately following seed imbibition, PEPC activity and concentration increase in parallel, with the greatest levels attained by the third day of germination. It is suggested that during this early phase of seed germination PEPC has a critical function to build up cellular dicarboxylic acid pools required to initiate significant activities of both the tricarboxylic acid and glyoxylate cycles.

Purification of two forms of starch branching enzyme (Q-enzyme) from developing rice endosperm.

Abstract: Two isoforms of starch branching enzyme (Q-enzyme), QEI and QEII, have been purified to honlogeneity from developing rice endosperm. QEI and QEII, with molecular weights of about 80 and 85 kDa, respectively, could be fully separated by anion-exchange or hydrophobic chromatography. The peptide maps obtained after V8 proteinase digestion were quite different between the two enzymes. Antibodies prepared against QEI showed no immunological cross-reaction with the QEII protein in Western blot experiments, and anti-QEII serum did not react with the QEI protein. The data indicate that QEI and QEII are distinct proteins encoded by different genes in rice plants.

Evidence of a pre-angiosperm origin of endosperm: implications for the evolution of flowering plants.

Abstract: The formation of a polyploid endosperm tissue has long been considered a unique and defining feature (autapomorphy) of angiosperms. Contemporaneous with the fertilization of an egg nucleus by a sperm nucleus in Ephedra trifurca (a nonflowering seed plant closely related to angiosperms), a second fertilization event has previously been shown to occur between a second sperm nucleus and the sister nucleus of the egg nucleus. Development of the second fertilization product is now shown to be fundamentally similar to that of endosperm in primitive flowering plants: both are characterized by an initial period of free nuclear proliferation followed by a process of cellularization. In Ephedra, however, the second fertilization product ultimately yields additional embryos. If double fertilization in Ephedra and angiosperms is evolutionarily homologous, it is likely that endosperm evolved from a supernumerary fertilization event that originally produced embryos into one that produced a specialized nonembryo tissue dedicated to the nourishment of the zygotic embryo.

Transport of nutrients into developing seeds: a review of physiological mechanisms

Abstract: After synthesis in the vegetative parts of the plant, assimilates are translocated to fruits through xylem and phloem. Research on factors controlling nutrient transport into developing seeds via the phloem has been stimulated by the development of the empty seed coat technique.There is a consensus that as assimilates are transported from maternal tissues to filial tissues they are delivered to the extracellular space (the apoplast) separating the two generations, prior to uptake from the apoplast into the tissues of the embryo or endosperm. The empty seed coat technique has been used for the study of several aspects of nutrient transport into seeds, e.g. metabolic control and turgorsensitive transport. The osmotic environment of seed tissues has a strong effect on assimilate transport into empty seeds. Several lines of evidence suggest that one of the main ‘secrets’ of the high sink strength of developing seeds, at least in many taxonomic groups of dicotyledons, is that the sink end of the phloem pathway is ‘bathed’ in an apoplast solution with a high concentration of osmotically active solutes. Data on maize do not fit this pattern. A turgor homeostat mechanism may help to maintain high solute concentrations in the seed apoplast. The apoplast environment of seed tissues may also stimulate synthesis of storage proteins and be involved in the prevention of precocious germination. In addition to the osmotic environment, other factors influencing sink strength are discussed. Some aspects of solute transformation during transport through seed tissues are described.

The importance of endosperm balance number in potato breeding and the evolution of tuber-bearing Solanum species

Abstract: Endosperm failure is considered the primary reason for the lack of success in intra-and interspecific crosses. The Endosperm Balance Number (EBN) hypothesis is a unifying concept for predicting endosperm function in intraspecific, interploidy, and interspecific crosses. In the EBN system, every species has an ‘effective ploidy’ (EBN), which must be in a 2:1 maternal to paternal ratio in the endosperm for crosses to succeed. The knowledge of EBN is very useful in the transfer of genes from exotic germplasm, and in the development of new breeding schemes in potato. The paper describes the strategies for introducing 2x(1EBN), 2x(2EBN), 4x(2EBN) and 6x(4EBN) germplasm into the cultivated 4x(4EBN) potato gene pool. A new methodology for producing 4x(4EBN) and 2x(2EBN) chromosome addition lines is also discussed. EBN has evolutionary importance in the origin of tuber-bearing Solanums. The role of the EBN in the origin of diploid and polyploid potato species, and as a barrier for hybridization and speciation of sympatric species within the same ploidy level is demonstrated. The origin of 3x and 5x cultivated tuber-bearing Solanums may also be explained using the EBN concept. EBN has been reported to exist in other plant species: alfalfa, beans, blueberries, rice, soybeans, squashes, tomato, forage legumes, grasses, ornamentals and Datura stramonium. This indicates that EBN may have broad application and could be useful for germplasm transfer and breeding in other crop species.

PCR amplification and sequences of cDNA clones for the small and large subunits of ADP-glucose pyrophosphorylase from barley tissues.

Abstract: Several cDNAs encoding the small and large subunit of ADP-glucose pyrophosphorylase (AGP) were isolated from total RNA of the starchy endosperm, roots and leaves of barley by polymerase chain reaction (PCR). Sets of degenerate oligonucleotide primers, based on previously published conserved amino acid sequences of plant AGP, were used for synthesis and amplification of the cDNAs. For either the endosperm, roots and leaves, the restriction analysis of PCR products (ca. 550 nucleotides each) has revealed heterogeneity, suggesting presence of three transcripts for AGP in the endosperm and roots, and up to two AGP transcripts in the leaf tissue. Based on the derived amino acid sequences, two clones from the endosperm, beps and bepl, were identified as coding for the small and large subunit of AGP, respectively, while a leaf transcript (blpl) encoded the putative large subunit of AGP. There was about 50% identity between the endosperm clones, and both of them were about 60% identical to the leaf cDNA. Northern blot analysis has indicated that beps and bepl are expressed in both the endosperm and roots, while blpl is detectable only in leaves. Application of the PCR technique in studies on gene structure and gene expression of plant AGP is discussed.

Histo-differentiation and molecular biology of developing cereal endosperm

Abstract: To elucidate the molecular aspects of cereal endosperm development, a good understanding of the histo-differentiation process is necessary. Reviewing the extensive history of lightand electron-microscopic investigations has revealed similarities between cereals with respect to cell ontogeny. Thus, using barley as a model and taking into account many developmental mutants in this and other species, a model for endosperm differentiation is proposed. Based on this model, molecular studies on endosperm gene-regulation are discussed and the available techniques for further investigations are considered.

A gibberellin-regulated gene from wheat with sequence homology to cathepsin B of mammalian cells.

Abstract: Summary A previous report described several cDNAs corresponding to mRNAs which accumulated in wheat aleurone layers treated with gibberellic acid (GA) (Baulcombe and Buffard, 1983). The protein sequence deduced from one of these clones (2529) has extensive similarity to the thiol protease, cathepsin B from mammalian cells. Southern analysis of wheat DNA has shown that the 2529 mRNA is encoded by a small family of genes carried on the group 4 chromosome. The nucleotide sequence of a member of the gene family expressed at a low level in aleurone layers and the use of a primer extension assay to identify a clone of a member of the gene family producing an abundant mRNA are reported. The 2529 mRNA accumulates in the scutellum and the aleurone layer of germinating grains where its expression is regulated by GA. In the scutellum the expression was restricted to the parenchyma, suggesting that the 2529 product may have a role other than for mobilization of the endosperm.

Effect of the floury-2 locus on protein body formation during maize endosperm development

Abstract: The seed storage proteins of maize (Zea mays L.) are synthesized during endosperm development on membrane-bound polyribosomes. These proteins, collectively called zeins, are translocated into the lumen of the rough endoplasmic reticulum, where they assemble into protein bodies. Protein body formation in normal genotypes occurs via an ordered deposition of the various types of zeins, and leads to the formation of spherical structures with a diameter of about 1 μm. These structures consist of a central core that contains predominantly α-zein; this central region is surrounded by a peripheral layer of β- and γ-zeins, and the entire structure is bounded by rough endoplasmic reticulum.

Genomic Nucleotide Sequence of a Wild-Type Shrunken-2 Allele of Zea mays

Abstract: The maize (Zea mays) endosperm enzyme, ADP-glucose pyrophosphorylase, which produces ADP-glucose from ATP and glucose-l-phosphate, is an important enzyme in the synthesis of starch. ADP-glucose arising from the action of this enzyme is the major, if not sole, donor of glucose for starch biosynthesis. The enzyme is composed of two dissimilar subunits encoded by the two unlinked genes, Shrunken-2 (Sh2) and Brittle-2 (Bt2) (1, 2). The enzyme is allosterically activated by 3-phosphoglyceric acid and inhibited by phosphate (3). Although it remains an open question whether these allosteric properties are physiologically relevant or whether they simply reflect the evolutionary history of the two structural genes (4), there does exist, nevertheless, much interest in determining whether genetic modification of this enzyme could lead to increased rates of starch biosynthesis in the maize endosperm. Because endosperm starch content comprises approximately 70% of the dry weight of the seed, alterations in starch biosynthesis would clearly affect corn yield. As a first step toward such experiments, we have isolated and sequenced genomic clones of Sh2. The structure of the gene (Table I, Fig. 1) is based on sequence analysis of three overlapping clones isolated from a Black Mexican Sweet genomic library. Exonic sequences were defined by comparison with the cDNA sequenced previously (2) and further sequencing of cDNAs subsequently isolated. These sequences correct for the fact that the original cDNA contained some non-Sh2 sequences at its extremities. The exonic genomic and cDNA sequences are nearly 100% identical. The few differences may be due to DNA polymorphisms because these clones were isolated from different corn lines. Placement of the start of the first exon is based on primer extension experiments. In the absence of dideoxynucleotides, four major bands, differing by one nucleotide, were observed. In the presence of dideoxynucleotides, the three large bands were seen in all four sequencing tracks. In each case, the largest band occurred at the same distance from the primer. It is unknown whether this reflects heterogeneity within the normal population of Sh2 transcripts or some laboratory artifact. Nevertheless, the start of transcription can be placed within three base pairs.

Galactomannan hydrolyzing activity develops during priming in the micropylar endosperm tip of tomato seeds

Abstract: Development of galactomannan hydrolyzing activity was followed in seeds of tomato [Lycopersicon esculentum (L..) Mill. cv. Toyonishiki] during priming and germination. The activity developed in seeds that were being primed in polyethylene glycol (-0.8 MPa). The activity was detected exclusively in the endosperm portion just adjacent to the radicle tip. Part of the activity remained active after desiccation of the primed seeds