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  • Author or Editor: Hong Xu x
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Five sequence characterized amplified region (SCAR) DNA markers were reevaluated at substantially higher annealing temperatures than originally reported; four were polymorphic among nine rootstocks tested. Four new informative SCAR markers also are reported, based on redesigning primers from previously cloned random amplified polymorphic DNA (RAPD) markers. Based on the eight polymorphic markers, rootstocks MG 420A, MG101-14, Richter 99, Couderc 3309, and Kober 5BB were distinguishable. Riparia Gloire and Couderc 1616 could be distinguished from the others, but not from one another, and SO4 and 5C also could be distinguished from the others, but not from one another.

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A total of eight random amplified polymorphic DNA (RAPD) markers were generated in a screen of 77 primers of 10-base length and were detected reproducibly among nine different grape (Vitis) rootstocks. Occasional failed amplifications could not be explained rationally nor easily corrected by systematic replacement of individual reaction components. In an effort to improve their reliability, the RAPD markers were cloned, their termini sequenced, and new sequence-specific primer pairs were synthesized based on addition of 10 to 14 bases to the 3' termini of the original 10-mers. Six pairs of the new primers were evaluated at their optimal and higher-than optimal annealing temperatures. One primer pair amplified a product the same size as the original RAPD marker in all rootstocks, resulting in loss of polymorphism. Post-amplification digestion with 7 different restriction endonucleases failed to reveal restriction site differences. Three primer pairs amplified an unexpected length variant in some accessions. Two other pairs of primers amplified a number of unexpected bands. Better approaches for exploiting the sequence differences that account for the RAPD phenomenon will be discussed.

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With the increase in dialysis patients worldwide, the demand for low-K vegetables is growing. Thus, a type of lettuce with a low-K content has been marketed in Japan. To learn more about low-K vegetables, information is needed on the physiological differences between these vegetables and those with typical levels of potassium (K). In this study, lettuces (Lactuca sativa L.) were cultivated using two low-K management methods in an environment-controlled system. One method was based on electrical conductivity (EC) management, and the K was replaced by sodium (Na) at the end of cultivation. The other method was based on quantitative nutrient management, and the nutrients required for low-K lettuce were quantitatively supplied, but no extra Na was added. Meanwhile, lettuce with normal K concentration was cultivated with EC management as the control. Plant growth indices, leaf photosynthesis traits, chlorophyll fluorescence characteristics, concentrations of secondary metabolites (SMs), and antioxidant activity were examined to investigate the physiological effects of low-K and high-Na concentrations during low-K lettuce cultivation. Both low-K treatments significantly restrained the growth of lettuce and increased the concentration of soluble sugar. However, photosynthesis and fluorescence characteristics remained unchanged. This indicates that the biomass reduction of low-K lettuce was due to the wasteful accumulation of carbohydrates rather than the decline in photosynthesis. Concentrations of SMs were increased in the low-K lettuce. In addition, higher concentrations of Na influenced the concentration of SMs, indicating that SMs were more sensitive to environmental stress.

Open Access

Randomly amplified polymorphic DNA (RAPD) markers were generated for identifying grape (Vitis) rootstocks. Seventy-seven primers (10 bases long) were screened using CsCl-purified leaf DNA derived from several field samples of nine rootstocks sampled in successive years. Nine RAPD markers were detected from six primers and, in combination, distinguished all nine rootstocks tested. Because inconsistencies were encountered in performing the RAPD assay, sequence-specific primers were derived from cloned RAPD bands for use under more stringent amplification conditions. Southern hybridization analysis of the RAPD gels with cloned RAPD bands as probes revealed deficiencies of scoring RAPD bands based solely on ethidium bromide staining. In some cases, bands of the same size generated by the same primer in different rootstocks-normally scored as the same marker-failed to cross-hybridize, implying lack of homology between the bands. More commonly, bands scored as absent based on ethidium bromide staining were detected by hybridization. Six of the nine cloned RAPD bands were partially sequenced, and sequence-specific primer pairs were synthesized. Two primer pairs amplified a product the same size as the original RAPD band in all rootstocks, resulting in loss of polymorphism. Two other pairs of sequence-specific primers derived from the same marker failed to amplify the expected band consistently. Three of the most useful primer pairs amplified apparent length variants in some accessions and will have value as polymerase chain-reaction markers for fingerprinting.

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Petal anthocyanins were systematically identified and characterized by high-performance liquid chromatography (HPLC)–electrospray ionization–mass spectrometry (MS) coupled with diode array detection among nine wild herbaceous peony (Paeonia L.) species (15 accessions). Individual anthocyanins were identified according to the HPLC retention time, elution order, MS fragmentation patterns, and by comparison with authentic standards and published data. Six main anthocyanins, including peonidin-3,5-di-O-glucoside, peonidin-3-O-glucoside-5-O-arabinoside (Pn3G5Ara), peonidin-3-O-glucoside, pelargonidin-3,5-di-O-glucoside, cyanidin-3,5-di-O-glucoside, and cyanidin-3-O-glucoside (Cy3G), were detected. In addition to the well-known major anthocyanins, some minor anthocyanins were identified in herbaceous peony species for the first time. Detection of the unique anthocyanins cyanidin-3-O-glucoside-5-O-galactoside and pelargonidin-3-O-glucoside-5-O-galactoside in both Paeonia anomala L. and P. anomala ssp. veitchii (Lynch) D.Y. Hong & K.Y. Pan indicated these two species should belong to the same taxon. Pn3G5Ara was found only in European wild species and subspecies suggesting different metabolic pathways between European and Chinese accessions. Anthocyanins conjugated with galactose and arabinose were observed in the genus Paeonia for the first time. The North American species, Paeonia tenuifolia L., had high Cy3G content in flower petals. This anthocyanin composition is distinct from the anthocyanin composition in Asian and European species and possibly is responsible for the vivid red coloration in flowers.

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A full-length cDNA isolated from banana (Musa acuminata L. AAA group) fruit was named MaMDH, containing an open reading frame encoding 332 amino acids that represents the gene for cytoplasmic malic dehydrogenase (MDH). Sequence analysis showed that MaMDH shares high similarity with MDHs from castor bean (XP_002533463), tobacco (CAC12826), peach (AAL11502), and chickpeas (CAC10208). Real-time quantitative polymerase chain reaction (PCR) analysis of MaMDH spatial expression showed that it was expressed in all organs examined: roots, rhizomes, leaves, flowers, and fruits. The expression was the highest in flowers followed by the fruits and roots, whereas the rhizomes and leaves displayed the lowest expression levels. Real-time quantitative PCR revealed that MaMDH exhibited differential expression patterns in post-harvest banana fruits correlating with ethylene biosynthesis. In naturally ripened banana fruits, MaMDH expression was in accordance with ethylene biosynthesis. In accordance, for banana fruits treated with the ethylene analog 1-methylclopropene (1-MCP), MaMDH expression levels were inhibited and remained constant. After treatment with ethylene, MaMDH expression in banana fruits significantly increased with ethylene biosynthesis and peaked 3 days after harvest, which was 11 days earlier than that in naturally ripened banana fruits. These results suggest that MaMDH expression is induced by ethylene to regulate post-harvest banana fruits ripening.

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The banana, a typical climacteric fruit, undergoes a postharvest ripening process followed by a burst in ethylene production that signals the beginning of the climacteric period. Postharvest ripening plays an important role in improving the quality of the fruit as well as limiting its shelf life. To investigate the role of glutamate decarboxylase (GAD) in climacteric ethylene biosynthesis and fruit ripening in postharvest banana, a GAD gene was isolated from banana, designated MuGAD. Coincidently with climacteric ethylene production, MuGAD expression as well as the expression of the genes encoding the Musa 1-aminocyclopropane-1-carboxylate synthase (MaACS1) and Musa 1-aminocyclopropane-1-carboxylate oxidase (MaACO1) greatly increased during natural ripening and in ethylene-treated banana. Moreover, ethylene biosynthesis, ripening progress, and MuGAD, MaACS1, and MaACO1 expression were enhanced by exogenous ethylene application and inhibited by 1-methylcyclopropene (1-MCP). Taken together, our results suggested that MuGAD is involved in the fruit ripening process in postharvest banana.

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Ferric chelate reductase (FRO) is a critical enzyme for iron absorption in strategy I plants, reducing Fe3+ to Fe2+. To identify FRO family genes in the local Citrus junos cultivar Ziyang Xiangcheng and to reveal their expression model, the citrus (Citrus sp.) genome was searched for homologies of the published sequence CjFRO1. Five FROs were found, including CjFRO1; these were named CjFRO2, CjFRO3, CjFRO4, and CjFRO5, respectively, and cloned via reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) PCR. The deduced amino acid sequences of five CjFROs contained flavin adenine dinucleotide (FAD)-binding motifs, nicotinamide adenine dinucleotide (NAD)-binding motifs, and 6–10 transmembrane domains, with isoelectric points between 6.73 and 9.46, and molecular weights between 67.2 and 79.9 kD. CjFRO1 and CjFRO2 were predominantly found in the aboveground parts of C. junos, with CjFRO1 highly expressed in leaves, and CjFRO2 largely expressed in stems and leaves. CjFRO3 was less expressed in roots, stems, and leaves. CjFRO4 and CjFRO5 were predominately found in roots. Under iron-deficient conditions, CjFRO4 was significantly and specifically increased in the roots of C. junos, whereas CjFRO1 was upregulated in the roots and leaves.

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