Objective: Information regarding the vitamin content of silage is limited. This study investigated the changes in the vitamin content of alfalfa and Chinese leymus silages with or without a lactic acid bacterial inoculant. Methods: Alfalfa at the early flowering stage and Chinese leymus at the full-bloom stage were harvested. The treatments for each forage type were control (deionized water only) and $1{\times}10^6$ colony-forming units Lactobacillus plantarum (LP)/g fresh matter. After 45 days of ensiling, all silages were sampled for evaluating the vitamin content, fermentation quality and chemical composition. Results: The LP inoculant decreased the pH value and ammonia nitrogen content of the alfalfa and Chinese leymus silages and significantly (p<0.05) increased the lactic acid, acetic acid concentrations and Flieg's points. Prior to ensiling, the levels of five B-group vitamins (thiamin, riboflavin, niacin, pantothenic acid, and pyridoxine) and ${\alpha}$-tocopherol in alfalfa were significantly (p<0.01) higher than those in Chinese leymus. Ensiling decreased the levels of the five B-group vitamins in both alfalfa and Chinese leymus while increasing the ${\alpha}$-tocopherol content of Chinese leymus. The thiamin, riboflavin, niacin and pantothenic acid levels in the LP-treated silage were significantly (p<0.05) lower than those in the untreated silage for the alfalfa and Chinese leymus. The ${\alpha}$-tocopherol content in the LP-treated alfalfa silage was significantly (p<0.05) higher than that in the untreated alfalfa silage. There was no significant (p>0.05) difference in pyridoxine content between the untreated and LP-treated silages for both forages. Conclusion: With or without LP inoculation, the levels of the five B-group vitamins (thiamin, riboflavin, niacin, pantothenic acid, and pyridoxine) in alfalfa and Chinese leymus decreased after 45 days of ensiling, while the ${\alpha}$-tocopherol content of Chinese leymus increased. The LP inoculant improved the fermentation quality of both the alfalfa and Chinese leymus silages but increased the thiamin, riboflavin, niacin, and pantothenic acid loss in the two forages after fermentation.
The objective of this study was to identify the effect of zebularine, a DNA methylation inhibitor, on the chromosomal association between homoeologous chromosomes in the wheat genetic background. Zebularine at a final concentration of 10 µM was used to treat the spikes of the double monosomic wheat addition line (DMA) with one Leymus mollis chromosome and one Leymus racemosus chromosome, both of which were in a homoeologous relationship. In late prophase, zebularine led to chromosome breakage in the Leymus homoeologous chromosomes. Chromosome breakage caused an increase in the frequency of chromosomal associations between the Leymus homoeologous chromosomes. Ordinary DMA showed 65 cells (35.3%) with chromosomal associations and 119 cells (64.7%) with no association, whereas treated DMA showed 102 cells (60.0%) with chromosomal associations and 67 cells (39.4%) with no association. In diakinesis, the Leymus bivalent showed a chromosomal association in the whole euchromatic region. In metaphase, the Leymus bivalent showed association in the whole chromosomal region, unlike other Leymus bivalents with partial chromosomal association. Chromosomal association by chromosome breakage occurred not only between Leymus chromosomes but also between Leymus and wheat chromosomes. The frequency of other chromosomal association (such as fusion and insert) was increased. Chromosome breakage by zebularine treatment is a useful method at the chromosome level as the spores with others are hereditary stable, although the homologous index (h) was not significantly different between ordinary DMA and treated DMA. It is necessary to study how to control zebularine treatment with a more stable concentration for chromosome breakage during meiosis.
The effect of rye B chromosome (rye B) on chromosome association was investigated in meiosis of wheat addition line. The wheat addition line was with one Leymus mollis chromosome and one L. racemosus chromosome which are under homoeologous relationship. Chromosome behavior of the two Leymus chromosomes in wheat genetic background was revealed by genomic in situ hybridization. In the first metaphase, most of the two Leymus chromosomes showed univalent in the wheat addition line without rye B (98.1 ± 0.5%). On the other hand, the wheat addition line with rye B showed higher frequency of bivalent (10.3 ± 0.2%) than wheat addition line without rye B (1.9 ± 0.5%). The wheat addition line without rye B showed abnormal bivalents with abnormal structure while the wheat addition line with rye B showed normal bivalent in low frequency. By rye B, some bivalent was composed of wheat and L. racemosus, and some trivalent was composed of wheat bivalents with L. mollis chromosome. Also, some wheat bivalents showed hyper-crossover, so those wheat bivalents showed abnormal structure compared to other wheat bivalents with normal structure such as ring, rod, and pan.
Lee, Won Ju;Cho, Seong-Woo;Tsujimoto, Hisashi;Roy, Swapan Kumar;Kim, Hong-Sig;Woo, Sun Hee
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.122-122
/
2017
Wheat wild relatives that have never been domesticated contained useful genetic resources such as the resistance to abiotic and biotic stresses. Leymus racemosus is one of the wild species. It can grow in a harsh environment like seaside and distribute by healthy rhizomes. Also, it has a useful genetic resource such as salt tolerance and different diseases resistance. Wheat (Triticum aestivum L. cv. Chinese Spring; CS) was crossed with L. racemosus. Wheat-L. racemosus disomic addition lines were produced. The purpose of this study is to identify protein expression in each disomic addition line compared to CS. We performed two-dimensional electrophoresis. Two-dimensional gels stained with coomassie brilliant blue (CBB), a total of 1566 differentially expressed proteins were identified by Progenesis Same Spots software from the cultivars. However, a total of 90 protein spots were identified to be either present or absent or showing significantly differential expression when the difference threshold was set to more than 1.5 fold. However, out of the 90 differentially protein spots, a total of 74 spots were sorted for mass spectrometry analysis. The identified proteins may provide important clues for better understanding the molecular changes in the chromosomes carrying Leymus racemosus.
The halophyte Leymus chinensis (Trin.) is a perennial rhizome grass (tribe Gramineae) that is widely distributed throughout China, Mongolia and Siberia. This study was conducted to investigate an optimal condition for plant regeneration from mature seeds, leaf base segments, and root segments in L. chinensis. Plant growth regulators affecting embryogenic callus induction and plant regeneration were investigated by four-factor-three-level [L9 (34)] orthogonal test in this study. The effects of explants types (mature seeds, leaf base segments and root segments), callus types, medium types were examined in this study. Wild type (WT) and Jisheng No. 1 plants (JS) were used for primary callus induction. A clear explants difference was seen during callus induction; mature seeds were considered as the preferred explants; and the highest frequency of callus induction was obtained in Medium 6 using mature seeds as explants in WT. Plant regeneration ability was evaluated by frequencies of green callus forming, shooting, rooting, and shooting with roots. Effect of α-naphthalene acetic acid (NAA) on shoot regeneration was remarkable with the highest frequency of 70.8% in WT after 2-month culture. The medium with 0.2- 0.5 mg/L NAA was found to have the highest shoot induction. All regenerated shoots were successfully rooted when transferred on half-strength Murashige and Skoog (MS) basal medium. The acclimatized plantlets were grown to mature with flowering and seeds setting in green house conditions.
To understand the adaptability of Leymus chinensis forage grass to environmental stresses, we analyzed the $H_2O_2$ scavenging activity based on several antioxidant enzymes and total phenolics content, including peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT), in shoots and roots subjected to salt and dehydration stresses during seedling growth. After NaCl or PEG treatment, plants showed reduced seedling growth under over 200 mM NaCl or 30% PEG treatment condition in shoots and roots compared with the control condition. In addition, plants showed high enzymatic activity of CAT in the shoots, whereas they exhibited high activity levels of APX and POD in the roots in both the NaCl and PEG treatment conditions. These results seem to indicate that Leymus chinensis seedlings responding to salt and dehydration stresses during initial growth is associated with enhanced activity of $H_2O_2$ scavenging antioxidant enzymes in the shoots or roots. The plants also showed high levels of total phenolics under NaCl treatment, with a high concentration in both the shoots and roots. Our results showed that the induced activity patterns of APX in the roots and CAT in the shoots indicate that low $H_2O_2$ levels were mainly maintained through tissue-specific redox homeostasis involving enzymes such as APX and CAT during salt and dehydration stresses. This study highlights the importance of antioxidant enzymes in the establishment of Leymus chinensis seedlings under high salinity conditions, such as typical desertification.
Kim Myoung Duck;Jin Hua;Park Eun-Joon;Kwon Suk-Yoon;Lee Haeng-Soon;Kwak Sang-Soo
Journal of Plant Biotechnology
/
v.32
no.1
/
pp.51-55
/
2005
Chinese leymus (Leymus chinensis Trin.) is a perennial grass that is widely distributed at high pH sodic and arid soil in the northeastern Asia. An efficient regeneration system was established through somatic embryogenesis of mature seeds to understand its high adaptability to harsh environmental conditions on the basis of molecular biology. The calli were efficiently induced (about $70\%$) from mature seeds on MS medium supplemented with $1.5\;\cal{mg/L}$ 2,4-D. Somatic embryos were formed from the surface of embryogenic callus on MS medium supplemented with $2.0\;\cal{mg/L}\;kinetin\;and\;0.5\;\cal{mg/L}$ NAA after 3 weeks of culture. Roots were induced from the shoot when transferred to MS medium without plant growth regulator for 1 week. Plant regeneration rate was $36\%$ and regenerated plantlets were grown to normal mature plants in pot. An efficient plant regeneration system in this study will be useful for molecular breeding of L. chinensis.
A study was conducted to investigate the seasonal changes in nutrient composition of pasture, voluntary intake and digestibility of sheep grazing an introduced Leymus chinensis pasture located in western Jilin Province, China. The whole-plant of L. chinensis and the samples simulating ingestion by sheep (simulating sample) were collected in spring (May, 2004), summer (July, 2004), autumn (September, 2004) and the end of winter (April, 2005). The contents of gross energy (GE), organic matter (OM), crude protein (CP), neutral detergent fibre (NDF), acid detergent fibre (ADF) and alkanes were determined. Voluntary intake and apparent digestibility of the nutrients in each season were also examined using 6 mature Chinese Northeast Merino ewes (differed among the seasons) grazing on a paddock of 1 ha size. The apparent digestibilities of GE, dry matter (DM), OM, CP, NDF and ADF of L. chinensis were significantly greater (p<0.05) in spring and summer than in autumn and winter. Metabolizable energy (ME) content was 12.13, 11.62, 8.66 and 10.40 MJ/kg DM for L. chinensis in spring, summer, autumn and winter respectively, and the corresponding voluntary feed intakes were 91, 119, 59 and 58 g/d per kg metabolic weight ($LW^{0.75}$). The ME contents and DM intakes in autumn and winter were significantly lower than in spring and summer (p<0.05). The intake of L. chinensis pasture was sufficient to provide ME requirements for maintenance by the dry ewes in all the seasons, but inadequate for maintenance protein requirement in winter. (Supported by funds from National Basic Research Program of China, Grant No. 2007CB106800).
Genomes of clusters of related eukaryotes are now being sequenced at an increasing rate. In this paper, we developed an accurate, low-cost method for annotation of gene prediction and exon-intron structure. The gene prediction was adapted for delta 1-pyrroline-5-carboxylate-synthetase (p5cs) gene from China wild-type of the halophytic Leymus chinensis (Trin.), naturally adapted to highly-alkali soils. Due to complex adaptive mechanisms in halophytes, more attentions are being paid on the regulatory elements of stress adaptation in halophytes. P5CS encodes delta 1-pyrroline-5-carboxylate-synthetase, a key regulatory enzyme involved in the biosynthesis of proline, that has direct correlation with proline accumulation in vivo and positive relationship with stress tolerance. Using analysis of reverse transcription-polymerase chain reaction (RT-PCR) and PCR, and direct sequencing, 1076 base pairs (bp) of cDNA in length and 2396 bp of genomic DNA in length were obtained from direct sequencing results. Through gene prediction and exon-intron structure verification, the full-length of cDNA sequence was divided into eight parts, with seven parts of intron insertion. The average lengths of determinated coding regions and non-coding regions were 154.17 bp and 188.57 bp, respectively. Nearly all splice sites displayed GT as the donor sites at the 5' end of intron region, and 71.43% displayed AG as the acceptor sites at the 3' end of intron region. We conclude that this method is a cost-effective way for obtaining an experimentally verified genome annotation.
The nutrient composition of pasture, voluntary intake and digestibility of diet ingested by goats grazing on an introduced Leymus chinensis pasture were measured across spring (May), summer (July), autumn (October) and winter (March). In each season, 12 Inner Mongolian Cashmere goats (6 wethers and 6 does with an average live weight of $22.2{\pm}1.3$ kg and $19.5{\pm}0.8$ kg, respectively) were used to graze on a 2 hectares size paddock. Diet selection was observed and the plant parts selected by grazing goats and whole plant L. chinensis were sampled simultaneously. The alkane pair $C_{32}:C_{33}$ and $C_{36}$ were used to estimate intake and digestibility, respectively. The results showed that the plant parts selected by goats had higher crude protein (CP) and lower acid detergent fiber (ADF) and neutral detergent fiber (NDF) than the whole plant, especially in the autumn and winter. The voluntary intake of dry matter (DM), CP, ADF, NDF, and metabolizable energy (ME) by goats was highest in summer (p<0.05). The goats ingested more CP, ME, and less ADF in spring than in autumn (p<0.05). The intakes of DM, CP, and ME were lowest in winter (p<0.05). There were significant differences in nutrient intake between wethers and does in each season, except for the ADF and ME intake per metabolic weight ($LW^{0.75}$). The nutrient digestibilities were higher in spring and summer, and decreased significantly during the autumn and winter (p<0.05). Goats, especially wethers, had a relative constant NDF digestibility across seasons, however, the apparent digestibility of CP in both wethers and does, decreased to negative values in winter. The grazing goats experienced relatively sufficient nutrients supply in spring and summer, and a severe deficiency of CP and ME in winter.
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