Quantification of carotenoids in citrus fruit by LC-MS and comparison of patterns of seasonal changes for carotenoids among citrus varieties

To quantify the 18 carotenoids on the basic routes of the carotenoid biosynthesis in plants simultaneously, a method for liquid chromatography-mass spectrometry (LC-MS) using atmospheric pressure chemical ionization was developed. With this method, the seasonal changes of carotenoids in the flavedo and juice sacs of 39 citrus varieties were analyzed. On the basis of the patterns of seasonal changes of carotenoids in both flavedo and juice sacs, 39 citrus varieties were classified. In flavedo, 39 varieties were classified into 5 clusters, in which the carotenoid profiles were carotenoid-poor, phytoene-abundant, violaxanthin-abundant, violaxanthin- and beta-cryptoxanthin-abundant, and phytoene-, violaxanthin-, and beta-cryptoxanthin-abundant, respectively. In juice sacs, they were classified into 4 clusters, in which the carotenoid profiles were carotenoid-poor, violaxanthin-abundant, violaxanthin- and phytoene-abundant, and violaxanthin-, phytoene-, and beta-cryptoxanthin-abundant, respectively. In flavedo, many citrus varieties, except for the carotenoid-poor and phytoene-abundant varieties, massively accumulated beta,epsilon-carotenoids (e.g., lutein), beta,beta-carotenoids (e.g., beta-cryptoxanthin and violaxanthin), and phytoene, in that order. In juice sacs, the accumulation order among beta,beta-carotenoids was observed. Violaxanthin accumulation preceded beta-cryptoxanthin accumulation in violaxanthin-, phytoene-, and beta-cryptoxanthin-abundant varieties. In each variety, the carotenoid profiles of the flavedo and juice sacs on the basis of the concentration in violaxanthin and beta-cryptoxanthin were similar, with the exception of a few varieties.     

Iron Uptake and Loading into Rice Grains

Iron (Fe) is an important micronutrient for living organisms. Fe deficiency severely impairs plant growth and is a widespread human dietary problem, with particularly high numbers of affected children and females. Rice (Oryza sativa) is a source of energy for more than half of the world’s population. Thus, understanding the mechanisms of Fe uptake and translocation in rice is of utmost importance in the development of rice varieties that are tolerant to low Fe availability and with high seed levels of Fe. In recent years, the mechanisms underlying Fe transport and homeostasis have been revealed, providing opportunities to increase the Fe content of rice grain. As excess Fe is toxic to cells, plants have developed sophisticated mechanisms to control Fe flow, making it difficult to alter Fe transport. Thus, choosing appropriate chelators and Fe transporters driven by appropriate promoters seems to be the key in developing rice that is tolerant to low Fe availability and which accumulates high grain levels of Fe. Many recent studies have been aimed at increasing the Fe content of rice. Here, we summarize these efforts and review recent progress in understanding the mechanisms of Fe transport.     

Structural characterization and hypoglycemic effects of arabinogalactan-protein from the tuberous cortex of the white-skinned sweet potato (Ipomoea batatas L.)

An arabinogalactan-protein (WSSP-AGP) was isolated from the tuberous cortex of the white-skinned sweet potato (WSSP; Ipomoea batatas L.). It consists of 95% (w/w) carbohydrate and 5% (w/w) protein with high contents of hydroxyproline, alanine, and serine. Its sugar composition is α-L-Rha:α-L-Ara:β-D-Gal:β-D-GlcA in a molar ratio of 1.0:4.1:7.6:1.3. Its weight-average molecular weight was estimated to be 126,800 g/mol by high-performance size exclusion chromatography coupled with multiangle laser light scattering. Structural analysis indicated that WSSP-AGP is a (1→3)-β-D-galactan highly branched at O-6 with (1→6)-β-D-galactan, in which the branched chains are substituted at the O-3 position with α-L-Araf-(1→ and α-L-Araf-(1→5)-α-L-Araf-(1→ and at the O-6 position typically with α-L-Rhap-(1→4)-β-D-GlcAp-(1→ as terminating groups. Continuous administration of WSSP-AGP to KKAy mice significantly lowered fasting plasma glucose levels. This indicates that WSSP-AGP plays an important role in the hypoglycemic effects of WSSP.     

High night temperature stimulates photosynthesis, biomass production and growth during the vegetative stage of rice plants

The effects of night temperature on biomass accumulation and plant morphology were examined in rice ( Oryza sativa L.) during vegetative growth. Plants were grown under three different night temperatures (17, 22 and 27°C) for 63 days. The day temperature was maintained at 27°C in all treatments. The final biomass of the plants was greatest in the plants grown at the highest night temperature. Total leaf area and tiller number were also the greatest in this treatment. Growth analysis indicated that the relative growth rate in the 27°C night-temperature treatment was maximal between days 21–42 and this was caused by increases in leaf area ratio, leaf weight ratio and specific leaf area. Plant total nitrogen contents did not differ among treatments. However, nitrogen allocation to the leaf blades was highest and the accumulation of sucrose and starch in the leaf blades and sheaths was the lowest in the 27°C night-temperature treatment by day 42. Despite this, dark respiration was also highest, and both the gross and net rates of CO₂ uptake at the level of the whole plant at day 63 were the highest in the 27°C night-temperature treatment. Thus, high night temperature strongly stimulated the growth of leaf blades during the early stage of rice plant growth, leading to increased biomass during the vegetative stage of the rice plants. As the CO₂ uptake rate per total leaf area was higher, photosynthesis at the level of the whole plant was also stimulated by a high night temperature.     

Analysis of the promoter of mutated human whey acidic protein (WAP) gene

Although whey acidic protein (WAP) has been identified in the milk of a range of species, it has been predicted that WAP is not secreted into human milk as a result of critical point mutations within the coding region. In the present study, we first investigated computationally the promoter region of mutated human WAP genes by comparing with those of other known WAP genes. Computational database analyses showed that the human WAP promoter region was highly conserved, as in other species with milk WAP. Next, we evaluated the activity of the human WAP promoter (2.6 kb) using a reporter gene assay. MCF-7 cells were stably transfected with the hWAP/hGH (human growth hormone) fusion gene, cultured on Matrigel, and treated with lactogenic hormones. Radioimmunoassay detected hGH in the culture medium, indicating that the human WAP promoter was responsible for the lactogenic hormones. The human WAP promoter was significantly more active in MCF-7 cells than the mouse WAP promoter (2.4 kb). The present results provide us with important information on the molecular evolution of milk protein genes.     

Effects of salt and low light intensity during the vegetative stage on susceptibility of rice to male sterility induced by chilling stress during the reproductive stage

We tested whether exposing rice plants to abiotic stress (salt or shade) during vegetative growth affects the chilling tolerance of reproductive organs, which is one of the most important traits for rice growing in a cool climate; we used two rice cultivars with different tolerance in two growing seasons. We divided the vegetative growth into three phases to clarify the most sensitive period: 7–22 days after transplanting (DAT), 23–38 DAT and 39–54 DAT. Chilling tolerance of the pre-stressed plants was based on the male sterility induced by low temperatures. Shade and salt stress during all three vegetative growth phases significantly reduced stomatal conductance. Shade decreased the specific leaf weight and the leaf sugar and starch contents, but salt had no significant effect, despite causing leaf damage. Low temperatures during the reproductive stage induced spikelet sterility in all plants, but the magnitude was greater in the salt- and shade-stressed plants of both cultivars, especially those stressed late during vegetative growth. The increased spikelet sterility caused by chilling was closely related to the reduction of the total spikelet number per panicle. This is the first study to show that salt and low light stress during vegetative growth increased the susceptibility of rice plants to chilling damage during panicle development.     

Molecular characteristics of porcine Fas-associated death domain (FADD) and procaspase-8

To reveal the intracellular signal transduction molecules involved in granulosa cell apoptosis in porcine ovarian follicles, we cloned the porcine Fas-associated death domain (FADD), an adaptor protein for the cell death receptor, and procaspase-8, an initiator caspase. Porcine FADD (pFADD) was 636 bp (211 amino acids: aa) long and showed 74.0 and 65.4% homology with human and murine FADD, respectively. Porcine procaspase-8 (pprocaspase-8) was 1,431 bp (476 aa) long and 70.6 and 63.4% homologous with human and murine procaspase-8, respectively. To confirm the apoptosis-inducing abilities, we constructed pFADD and pprocaspase-8 cDNA expression vectors with enhanced green fluorescence protein (EGFP) and then transfected them into human uterine cervix tumor (HeLa-K), human granulosa cell-derived (KGN), murine granulosa-derived tumor (KK1), and porcine granulosa cell-derived (JC410) cells. When pFADD and pprocaspase-8 were overexpressed, cell death was induced in these transfected cells. However when caspase-inhibitor p35 was cotransfected, cell death was inhibited. The pFADD and pprocaspase-8 genes are well conserved, as are the physiological functions of their products.