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Chromosome microdissection by laser microbeam, chromosomal fragment isolation and amplification in vitro in barley (hordeum vulgare L.)

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Chinese Science Bulletin 1998年第10期43卷 851-855页

作者： Huai Wang Yihua Zhou Benyuan Dang Zanmin Hu Lanlan Wang Liangcai Li Zhenghua Chen Acad Sinica Inst Genet Beijing 100101 Peoples R China

A method for microdissection, isolation and amplification of plant chromosomal fragments using laser microbeam and a glass microneedle was established. Firstly, 7H chromosome of barley (hordeum vulgare L.) was dissected by Nd: YAG laserbeam with suitable parameters and the fragment comprising a satellite was isolated with a glass microneedle which was fixed on a micromanipulator. Then, the chromosomal fragment DNA was amplified by LA_PCR (linker adaptor PCR) for two rounds. The size of the DNA fragments of PCR products varied from 500-3 000 bp and the PCR products originated from the genome of barley were verified by Southern hybridization. Compared with previous reports, there are some advantages in this research. The performance is easier, the dissection is more precise and the cost is low. It also permits efficient amplification with only one single chromosome fragment. Laser microbeam_glass microneedle method may be useful in the microdissection of special chromosome regions, especially in plants with middle or small chromosomes.

关键词： hordeum vulgare L. chromosome microdissection laser microbeam LA_PCR.

The effect of photo-irradiation on the growth and ingredient composition of young green barley (hordeum vulgare)

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Agricultural Sciences 2013年第4期4卷 185-194页

作者： Ryota Koga Tianxiao Meng Eriko Nakamura Chizuru Miura Nobuto Irino Hari Prasad Devkota Shoji Yahara Ryuichiro Kondo Department of Agro-Environmental Science Faculty of Agriculture Kyushu University Fukuoka Japan Graduate School of Pharmaceutical Sciences Kumamoto University Kumamoto Japan

We clarified that photo-irradiation of young green barley from three different light sources, natural light, 100% red light-emitting diodes (R-LEDs), and a mixture of 90% red-LEDs + 10% blue-LEDs (RB-LEDs), had significantly different results in growth degree (weight and height) and in components of young green barley. Barley that has sprouted for 15 days after germination did not show any apparent difference in height in response to irradiation by the three tested light sources, but by the 20th day of sprouting the height showed a positive effect by R-LEDs irradiation. By 15 days of sprouting the barley had achieved the heaviest weight by natural light irradiation, while the barley irradiated by R-LEDs had made remarkable progress at 20 days of sprouting. On the other hand, the irradiation by RB-LEDs showed a suppressive tendency after 15 days or more. The amino acid content, as indicated by dry weight conversion, was greatest in the barley irradiated by RB-LEDs, followed by R-LEDs, and natural light, which showed that LEDs irradiation is effective. In addition, four cyanogenic glucosides were isolated, identified, and quantified, as they are components frequently assessed in barley research. With regard to vitamin E, R-LEDs irradiation increased γ-tocopherol. Our results indicate that irradiation by LEDs would be effective for the enhancement of the functionality of young green barley.

关键词： hordeum Vulgare LEDs Cyanogenic Glucosides Amino Acid Tocopherol SEM

Model Examination for the Effect of Treading Stress on Young Green Barley (hordeum vulgare)

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American Journal of Plant Sciences 2013年第1期4卷 174-181页

作者： Ryota Koga Tianxiao Meng Eriko Nakamura Chizuru Miura Nobuto Irino Shoji Yahara Ryuichiro Kondo Department of Agro-Environmental Science Faculty of Agriculture Kyushu University Fukuoka Japan Graduate School of Pharmaceutical Sciences Kumamoto University Kumamoto Japan

Treading barley (“Mugifumi” in Japanese) is a barley culturing process that is unique and indispensable in Japan. Generally, roller machines or human feet are used to tread young barley several times (Figure 1). As a model examination, a stainless steel instrument was used for treading barley in this study (Figure 2(a)). Treading damages plants and decreases growth, such as the height and weight of aerial parts, which then recover gradually. In our study, the total amino acid contents increased with the treading stress. The treading stress induced a 1.7-fold increase in aspartic acid contents and an approximately 1.6-fold increase in glutamic acid, proline, cystine, and methionine contents. Isolation and purification suggested that the main components of the methanol-eluted fraction from the young green barley were six known phenolic compounds. Saponarin and lutonarin were the main components. The saponarin content was about 160 mg/g (methanol-soluble fraction) at 10 days after germination. We found that the higher antioxidant activity was due to the increased lutonarin/saponarin ratio from 10% to 24%. Therefore, our results suggest that treading stress is useful for young green barley.

关键词： hordeum vulgare Treading Stress Mugifumi Flavonoid Amino Acid Antioxidant Activity

Differential Responses of Antioxidative System to Soil Water Shortage in Barley (hordeum vulgareL.) Genotypes

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Advances in Biological Chemistry 2014年第6期4卷 351-359页

作者： Irada Mammad Huseynova Masma Yagubali Nasrullayeva Samira Mahammadrahim Rustamova Durna Rafail Aliyeva Jalal Alirza Aliyev Institute of Botany Azerbaijan National Academy of Sciences Baku Azerbaijan Institute of Genetic Resourses Azerbaijan National Academy of Sciences Baku Azerbaijan

Drought is one of the major factors limiting the yield and quality of crops in the world. The activity of antioxidative system to tolerate the drought stress is significant in plants. In the present study, the activities and isoform profiles of catalase (CАТ), ascorbate peroxidase (APX), glutathione reductase (GR), and superoxide dismutase (SOD) were analyzed in four barley genotypes grown under soil water restriction. Drought stress caused increase in the activities of CАТ and SOD in all studied genotypes, while APX activity decreased. The total GR activity increased substantially in genotypes K 2778 and *** 7 and decreased in No. 77 local and *** 596 genotypes under conditions of severe water stress. No detectable differences were observed in the isoenzyme pattern (the appearance of a new isoenzymes and disappearance of another one) between control plants and those subjected to soil drought. However, intensification of corresponding isoforms in electrophoretic spectra was observed in stressed barley leaves relative to watered ones. The obtained results possibly suggest that antioxidant protection in barley plants under drought conditions could be attributed mainly to SOD and CAT.

关键词： hordeum vulgare L. Drought Reactive Oxygen Species Antioxidant Enzymes

Further molecular evidence for the hordeum vulgare ssp. spontaneum in Tibet as ultimate progenitor of Chinese cultivated barley

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High Technology Letters 2005年第3期11卷 320-324页

作者： Feng Zongyun（冯宗云） Zhang Lili Zhang Yizheng Ling Hongqing State Key Laboratory of Plant Cell ＆ Chromosome Engineering Institute of Gengties ＆ Developmental Biology Chinese Aeademy of Seiences Beijing 100101 P. R. China Key Laboratory of Molecular Biology ＆ Biotechnology College of Life Science Sichuan University Chengdu 610065 P. R. China College of Agronomy Sichuan Agricultural University Ya＇an 625014 P. R. China

106 aeeessions of Tibetan wild barley, including 50 accessions of the two-rowed wiht barley hordeum vulgare ssp. sopntaneum （HS）, 27 accessions of the six-rowed bottle-shaped wild barley H. lagunculiforme （HL） and 29 accessions of the six-rowed wiht barley H. agriocrithon （HA） that separately represent different agrigeographical regions of Tibet, were used to study the genetic diversity and genetie differentiation using SSR markers selected from seven barley linkage groups. 229 allelic variants were identified with an average of 7.6 alleles/locus. The average of total number of alleles per locus in HA （6.4） is much higher than that in HS （3.9） and HL （3.4）. The genetie diversity and its standard deviation among the three subspecies were in the order of HS〉HL〉HA. Very significant genetic differentiation was observed among the three subspecies of wild barley. Comparisons of the results fiom this and previous studies showed a strong Oriental-Occidental differentiation of barley, and that Shannan region of Tibet might be the center of origin of the Tibetan two-rowed wild barley, thus supporting not only the hypothesis of a mono-phyletie origin of cultivated barley but also the proposition that the Tibetan two-rowed wild barley as ultimate progenitor of Chinese cultivated barley.

关键词： hordeum genetic diversity SSR markers genetic differentiation wild barley

栽培大麦×野生大麦(hordeum brenisubula)的杂交研究(Ⅰ)——通过幼胚的离体培养获得远缘杂种植株

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延边大学农学学报 1989年第1期16卷 103-107,121页

作者：黄永秀刘宪虎赵诚坤荆绍凌延边农学院农学系作物育种教研室延边农学院农学系遗传教研室延边农学院农学系83级

本文对栽培大麦×野生大麦杂种胚的离体培养进行了初步研究,旨在为将野生大麦(hordeum brenisubula)的一些优良特性转移到栽培大麦提供有效的方法和途径。1986～1987年对7个栽培大麦×野生大麦组合的杂种胚进行了离体培养试验,结果如下... 详细信息

本文对栽培大麦×野生大麦杂种胚的离体培养进行了初步研究,旨在为将野生大麦(hordeum brenisubula)的一些优良特性转移到栽培大麦提供有效的方法和途径。1986～1987年对7个栽培大麦×野生大麦组合的杂种胚进行了离体培养试验,结果如下:对照试验(只杂交,不进行杂种胚的离体培养)的483个颖花无一结实,说明不进行离体培养杂种胚很难发育成种子。在离体培养的2206个杂种胚中共得到了91株绿苗。可以看出:由于杂交组合的不同(即母本不同),出苗率有显著差异,7个组合的平均出苗率为4.13%,变幅为1.72～12.71%,不同组合获得有效胚(可接种的胚)的成功率不同,认为选择适当的母本可以提高远缘杂交的成功率和出苗率;胚的大小不同,出苗率亦不同,大胚的平均出苗率为22.01%,变幅为2.78～43.75%,小胚的平均出苗率为1.65%,变幅为0.60～7.84%,大胚的出苗率显著高于小胚,因此认为提高大胚的比例是提高出苗率的有效途径;不同胚龄(授粉接种的天数)出苗率不同,授粉后12—19天接种的胚均出了苗,而20天以上的胚均未出苗,调查发现,胚龄为15天以上时,有效胚的百分率逐渐下降,认为接种的最佳时期为授粉后12～14天。91株绿苗经移栽成活了57株,根据形态可将57株成活株分成三种类型,即:父本(野生)类型、中间类型母本(栽培)类型。对三种类型进行了形态上的定性描述,并讨论了它们的形成机理。

关键词：离体培养大麦品种 hordeum 远缘杂种种胚幼胚球茎大麦颖花数中间类型胚龄

木石港3号大麦(hordeum vulgare L)抗黄花叶病和酯酶同功酶两个位点的连锁关系

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大麦与谷类科学 1991年第3期21卷 49-,28页

作者：小西猛朗河田尚之吉田久早乙女和彦蔡立旺

作者旨在确定木石港3号抗黄花叶病位点Ym和酯酶同功酶复合位点（Est1—Est2—Est4）的连锁关系,并且通过苗期酯酶同功酶电泳法,评估选择含Y基因的单株和品系的可能性。

关键词：连锁关系黄花叶同功酶复合位点 hordeum 石港杂合染色体长臂电泳法纯合体

栽培大麦×野生大麦(hordeum brevisubulatum)的杂交研究(Ⅱ)——种间杂种的胚胎学观察

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延边大学农学学报 1990年第2期17卷 1-5页

作者：刘宪虎黄永秀柳参奎赵诚坤许明子吴基日延边农学院农学系育种教研室延边农学院农学系遗专教研室

对栽培大麦×野生大麦杂交的胚胎发育情况进行了观察,结果如下:栽培大麦与野生大麦的种间杂交可交配性很差,多数子房均未受精,少数子房中观察到虽然能受精,但极不正常,受精延迟而且不完整,仅发生卵的单受精,并且发育中途停止,在众多制... 详细信息

对栽培大麦×野生大麦杂交的胚胎发育情况进行了观察,结果如下:栽培大麦与野生大麦的种间杂交可交配性很差,多数子房均未受精,少数子房中观察到虽然能受精,但极不正常,受精延迟而且不完整,仅发生卵的单受精,并且发育中途停止,在众多制片中末发现有胚乳发育,也未发现胚的进一步分化和成熟。说明栽培大麦与野生大麦杂交存在合子前和合子后面的困难,讨论了这些困难产生的可能原因,并认为引起合子后困难的主要原因可能是因为不能形成胚乳,因此认为通过杂种胚的离体培养可在一定程度上克服合子后发育困难,而能获得杂种植株。

关键词：大麦品种 hordeum 可交配性胚乳发育短芒大麦草单受精种间杂种离体培养胚胎发育情况种胚

雅鲁藏布江中部流域发现古青稞（hordeum vulgare ***． nudum）炭化粒

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西南大学学报(自然科学版) 1994年第6期 576-576页

作者：傅大雄张俊卿田存余巴果次仁西藏自治区贡嘎县农业局西南农业大学农学及生命科学院麦类研究室西藏自治区贡嘎县水电局

雅鲁藏布江中部流域发现古青稞（ＨｏｒｄｅｕｍｖｕｌｇａｒｅＬ．ｖａｒ．ｎｖｄｕｍ）炭化粒傅大雄（西南农业大学农学及生命科学院麦类研究室）张俊卿，田存余，巴果，次仁（西藏自治区贡嘎县农业局，西藏自治区贡嘎县水电局）１９... 详细信息

雅鲁藏布江中部流域发现古青稞（ＨｏｒｄｅｕｍｖｕｌｇａｒｅＬ．ｖａｒ．ｎｖｄｕｍ）炭化粒傅大雄（西南农业大学农学及生命科学院麦类研究室）张俊卿，田存余，巴果，次仁（西藏自治区贡嘎县农业局，西藏自治区贡嘎县水电局）１９９４年９月６日，作者一行在地处雅鲁...

关键词：巴果青稞元麦贡嘎县雅鲁藏布江 nudum var vulgare hordeum 流域