Expression and parent-of-origin effects for FIS2, MEA, and FIE in the endosperm and embryo of developing Arabidopsis seeds

The promoters of MEA (FIS1), FIS2, and FIE (FIS3), genes that repress seed development in the absence of pollination, were fused to β-glucuronidase (GUS) to study their activity pattern. The FIS2∷GUS product is found in the embryo sac, in each of the polar cell nuclei, and in the central cell nucleus. After pollination, the maternally derived FIS2∷GUS protein occurs in the nuclei of the cenocytic endosperm. Before cellularization of the endosperm, activity is terminated in the micropylar and central nuclei of the endosperm and subsequently in the nuclei of the chalazal cyst. MEA∷GUS has a pattern of activity similar to that of FIS2∷GUS, but FIE∷GUS protein is found in many tissues, including the prepollination embryo sac, and in embryo and endosperm postpollination. The similarity in mutant phenotypes; the activity of FIE, MEA, and FIS2 in the same cells in the embryo sac; and the fact that MEA and FIE proteins interact in a yeast two-hybrid system suggest that these proteins operate in the same system of control of seed development. Maternal and not paternal FIS2∷GUS, MEA∷GUS, and FIE∷GUS show activity in early endosperm, so these genes may be imprinted. When fis2, mea, and fie mutants are pollinated, seed development is arrested at the heart embryo stage. The seed arrest of mea and fis2 is avoided when they are fertilized by a low methylation parent. The wild-type alleles of MEA or FIS2 are not required. The parent-of-origin-determined differential activity of MEA, FIS2, and FIE is not dependent on DNA methylation, but methylation does control some gene(s) that have key roles in seed development.

Influence of Water Content and Temperature on Molecular Mobility and Intracellular Glasses in Seeds and Pollen1

Although the occurrence of intracellular glasses in seeds and pollen has been established, physical properties such as rotational correlation times and viscosity have not been studied extensively. Using electron paramagnetic resonance spectroscopy, we examined changes in the molecular mobility of the hydrophilic nitroxide spin probe 3-carboxy-proxyl during melting of intracellular glasses in axes of pea (Pisum sativum L.) seeds and cattail (Typha latifolia L.) pollen. The rotational correlation time of the spin probe in intracellular glasses of both organisms was approximately 10−3 s. Using the distance between the outer extrema of the electron paramagnetic resonance spectrum (2Azz) as a measure of molecular mobility, we found a sharp increase in mobility at a definite temperature during heating. This temperature increased with decreasing water content of the samples. Differential scanning calorimetry data on these samples indicated that this sharp increase corresponded to melting of the glassy matrix. Molecular mobility was found to be inversely correlated with storage stability. With decreasing water content, the molecular mobility reached a minimum, and increased again at very low water content. Minimum mobility and maximum storage stability occurred at a similar water content. This correlation suggests that storage stability might be at least partially controlled by molecular mobility. At low temperatures, when storage longevity cannot be determined on a realistic time scale, 2Azz measurements can provide an estimate of the optimum storage conditions.

cis-Isomers of Cytokinins Predominate in Chickpea Seeds throughout Their Development1

Trans-isomers of cytokinins (CK) are thought to predominate and have greater biological activity than corresponding cis-isomers in higher plants. However, this study demonstrates a system within which the predominant CK are cis-isomers. CK were measured at four developmental stages in developing chickpea (Cicer arietinum L. cultivar Kaniva) seeds by gas chromatography-mass spectrometry. Concentrations were highest at an early endospermic fluid stage and fell considerably when the cotyledons expanded. The cis-isomers of zeatin nucleotide ([9R-MP]Z), zeatin riboside ([9R]Z), and zeatin (Z) were present in greater concentrations than those of corresponding trans-isomers: (trans)[9R-MP]Z, (trans)[9R]Z, (trans)Z, or dihydrozeatin riboside. Dihydrozeatin, dihydrozeatin nucleotide, and the isopentenyl-type CK concentrations were either low or not detectable. Root xylem exudates also contained predominantly cis-isomers of [9R-MP]Z and [9R]Z. Identities of (cis)[9R]Z and (cis)Z were confirmed by comparison of ion ratios and retention indices, and a full spectrum was obtained for (cis)[9R]Z. Tissues were extracted under conditions that minimized the possibility of RNase hydrolysis of tRNA following tissue disruption, being a significant source of the cis-CK. Since no isomerization of (trans)[2H]CK internal standards occurred, it is unlikely that the cis-CK resulted from enzymic or nonenzymic isomerization during extraction. Although quantities of total CK varied, similar CK profiles were found among three different chickpea cultivars and between adequately watered and water-stressed plants. Developing chickpea seeds will be a useful system for investigating the activity of cis-CK or determining the origin and metabolism of free CK.

Galactosylononitol and Stachyose Synthesis in Seeds of Adzuki Bean1 : Purification and Characterization of Stachyose Synthase

Stachyose synthase (STS) (EC 2.4.1.67) was purified to homogeneity from mature seeds of adzuki bean (Vigna angularis). Electrophoresis under denaturing conditions revealed a single polypeptide of 90 kD. Size-exclusion chromatography of the purified enzyme yielded two activity peaks with apparent molecular masses of 110 and 283 kD. By isoelectric focusing and chromatofocusing the protein was separated into several active forms with isoelectric point values between pH 4.7 and 5.0. Purified STS catalyzed the transfer of the galactosyl group from galactinol to raffinose and myo-inositol. Additionally, the enzyme catalyzed the galactinol-dependent synthesis of galactosylononitol from d-ononitol. The synthesis of a galactosylcyclitol by STS is a new oberservation. Mutual competitive inhibition was observed when the enzyme was incubated with both substrates (raffinose and ononitol) simultaneously. Galactosylononitol could also substitute for galactinol in the synthesis of stachyose from raffinose. Although galactosylononitol was the less-efficient donor, the Michaelis constant value for raffinose was lower in the presence of galactosylononitol (13.2 mm) compared with that obtained in the presence of galactinol (38.6 mm). Our results indicate that STS catalyzes the biosynthesis of galactosylononitol, but may also mediate a redistribution of galactosyl residues from galactosylononitol to stachyose.

Purification and Characterization of a Low-Molecular-Weight Phospholipase A2 from Developing Seeds of Elm1

Phospholipase A2 (PLA2) was purified about 180,000 times compared with the starting soluble-protein extract from developing elm (Ulmus glabra) seeds. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified fraction showed a single protein band with a mobility that corresponded to 15 kD, from which activity could be recovered. When analyzed by matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry, the enzyme had a deduced mass of 13,900 D. A 53-amino acid-long N-terminal sequence was determined and aligned with other sequences, giving 62% identity to the deduced amino acid sequence of some rice (Oryza sativa) expressed sequence tag clones. The purified enzyme had an alkaline pH optimum and required Ca2+ for activity. It was unusually stable with regard to heat, acidity, and organic solvents but was sensitive to disulfide bond-reducing agents. The enzyme is a true PLA2, neither hydrolyzing the sn-1 position of phosphatidylcholine nor having any activity toward lysophosphatidylcholine or diacylglycerol. The biochemical data and amino acid sequence alignments indicate that the enzyme is related to the well-characterized family of animal secretory PLA2s and, to our knowledge, is the first plant enzyme of this type to be described.

Xyloglucan Octasaccharide XXLGol Derived from the Seeds of Hymenaea courbaril Acts as a Signaling Molecule1

Treatment of the xyloglucan isolated from the seeds of Hymenaea courbaril with Humicola insolens endo-1,4-β-d-glucanase I produced xyloglucan oligosaccharides, which were then isolated and characterized. The two most abundant compounds were the heptasaccharide (XXXG) and the octasaccharide (XXLG), which were examined by reference to the biological activity of other structurally related xyloglucan compounds. The reduced oligomer (XXLGol) was shown to promote growth of wheat (Triticum aestivum) coleoptiles independently of the presence of 2,4-dichlorophenoxyacetic acid (2,4-D). In the presence of 2,4-D, XXLGol at nanomolar concentrations increased the auxin-induced response. It was found that XXLGol is a signaling molecule, since it has the ability to induce, at nanomolar concentrations, a rapid increase in an α-l-fucosidase response in suspended cells or protoplasts of Rubus fruticosus L. and to modulate 2,4-D or gibberellic acid-induced α-l-fucosidase.

Identification of Inositol 1,3,4-Trisphosphate 5-Kinase and Inositol 1,3,4,5-Tetrakisphosphate 6-Kinase in Immature Soybean Seeds

In extracts of immature soybean (Glycine max [L.] Merr.) seeds inositol tetrakisphosphate was formed from [3H]inositol 1,3,4-trisphosphate but not from [3H]inositol 1,4,5-trisphosphate. Inositol 1,3,4-trisphosphate kinase was purified to a specific activity of 3.55 min−1 mg−1 by polyethylenimine clarification and anion-exchange chromatography. The partially purified enzyme converted [3H]inositol 1,3,4-trisphosphate to inositol 1,3,4,5-tetrakisphosphate as the major product and inositol 1,3,4,6- and/or 1,2,3,4-tetrakisphosphate as the minor product. Subsequent experiments revealed a separate inositol 1,3,4,5-tetrakisphosphate 6-kinase activity, which could link these enzymes to inositol hexakisphosphate synthesis via the previously reported inositol 1,3,4,5,6-pentakisphosphate 2-kinase. The apparent Km values for inositol 1,3,4-trisphosphate kinase were 200 ± 0 nm for inositol 1,3,4-trisphosphate and 171 ± 4 μm for ATP, and the reaction was not reversible. The kinetics were such that no activity could be detected using unlabeled inositol 1,3,4-trisphosphate and [γ-32P]ATP, which suggested that other kinases may have been observed when less purified fractions were incubated with radiolabeled ATP. Inositol 1,3,4-trisphosphate kinase was nonspecifically inhibited more than 80% by various inositol polyphosphates at a concentration of 100 μm.

Inventory of seeds and plants imported / U.S. Department of Agriculture, Bureau of Plant Industry.

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Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 2

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet II [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng and southern Gansu Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 3

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet III [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng and southern Gansu Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 4

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet IV [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 9

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet IX [of 10] covering a portion of the Yellow River (Huang He) region in southern Gansu Sheng and northwestern Sichuan Sheng, China, including parts of Baishui Jiang and Pai Ho (Gar He). The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 5

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet V [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng and southern Gansu Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 6

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet VI [of 10] covering a portion of the Yellow River (Huang He) region in eastern Qinghai Sheng, southern Gansu Sheng, and northwestern Sichuan Sheng, China. The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.

Rock map of the Yellow River (Huang He) expedition, China, 1925-1927 : Sheet 7

This layer is part of a set of georeferenced, raster images of the manuscript, paper map set entitled: Ch'ing-Hai upper Yellow River expedition : Rock and Simpson, 1925-27, [cartography by J.F. Rock]. Scale 1:250,000. This layer image is of Sheet VII [of 10] covering a portion of the Yellow River (Huang He) region in southern Gansu Sheng and northwestern Sichuan Sheng, China, including parts of Bailong Jiang and Tao He. Sheet VII includes a separate title: 'Cho-ni Territory, Upper and Lower T'ieh-Pu country and route to Sung-P'an, J. F. Rock, 1925-1927.' The map set details the route and surrounding environs of the Arnold Arboretum's "Western China" expedition led by Joseph Rock, 1924-1927. The set covers a portion of the Yellow River (Huang He) region in south central China (Qinghai, Gansu, and Sichuan shengs (a portion of historic Tibet)). It shows features, labeled variously in English, Chinese, Wade-Giles transliteration, and Tibetan, including: rivers, streams, lakes, mountains, gorges, valleys, plateaus, plains, cities, towns, villages, provincial capitals, county seats, passes, monasteries, ruin sites, native tribe locations, and more. Relief is shown by hachures, spot heights, and landform drawings. The original manuscript map set is part of the Harvard College Library, Harvard Map Collection. "Joseph Rock traced his travels for the [Arnold] Arboretum's [Western China] 1924-1927 expedition in a colorful, hand-drawn map entitled 'Ch'ing-Hai upper Yellow River expedition.' The pen-and-ink drawing was made on ten sheets that when joined form a single, irregularly-shaped map, approximately six by eight feet in size. The individual sheets are numbered, using roman numerals; on sheet VII is a second title, 'Choni Territory, Upper and Lower T'ieh-Pu country and route to Sung-Pan, J. F. Rock, 1925-1927.' Topographical and other features are identified using a combination of English, Chinese characters, Wade-Giles transliterations and Tibetan script. Rock's attractive cursive style and use of hachures, spot heights, and landform drawings to depict relief add character to the map." -- Text from the Arnold Arboretum Web site.