Alfalfa weevil (Hypera postica Gyllenhal) belongs to the family Curculionidae, Order Coleoptera and it is one of invasive alien species in Korea. In Korea environment, this species was found in 2000 first time through the Korean quarantine inspections. Although various researches have been studied in the Europe, US, China and Japan, only few research has been performed in Korea. In this study, the life cycle, morphology and the host plants of Alfalfa weevil were investigated in the natural environment. Nine plants of total 28 host plant species known as the host plants were characterized as their wild host plants in Korea (6 and 3 of 9 were naturalized and native plants in Korea, respectively). The geographical distribution was very wide from south (Jeju) to north (Yeoncheon) area of the peninsula and the favorite habitats were park, cultivated and grass land. This species has been found easily anywhere because it takes common leguminous plants such as clover, chinese milk vetch and alfalfa. These results suggest that Alfalfa Weevil can be one of the most-damaging insect pests of leguminous crops when it adapts to Korean climate.

Alfalfa weevil is an invasive pest of several leguminous plants. In the world preservative, there is a lack of information about the level of differentiation among separate populations. We ran a comparative molecular and ecological analysis to assess diversity, phylogenetic relationship and also simplify the taxonomic decisions. By analyzing a total of 147 specimens from Middle East, Europe and America plus a few sequences in GenBank we improved our current knowledge of world phylogeography of Hypera postica. The result of mitochondrial genes (Summation of COI and CytB) revealed a deep divergence between two previously defined clades (5.6 – 6.7% nucleotide divergence) within sympatric distribution. The Eastern lineage comprised 60 haplotypes in Iran and 3 haplotypes in Italy with respectively 0.07 – 2.2% and 0.2 – 3.4% sequence divergence range. On the other hand, the Western lineage which was detected in Poland, Czech and Western America had relatively lower haplotype diversity presenting a total of 11 haplotypes and 0.1%-1.1% sequence divergence. The coexistence of two lineages in some regions led to the huge overlap in their fundamental nich, but also a certain level of divergence can be detected via ecological nich modeling analysis.

The present research investigated copper and cadmium stress-induced differentially expressed genes (DEGs) using annealing control primers (ACP) with the differential display reverse transcription polymerase chain reaction technique in alfalfa (Medicago sativa L. cv. Vernal) leaves. Alfalfa leaves were subjected to 250 μM of copper and cadmium treatment for a period of 6 h. A total of 120 ACPs was used. During copper and cadmium treatment, 6 DEGs were found to be up or down regulated. During copper stress treatment, 1 DEG was up-regulated, and 3 novel genes were discovered. Similarly, during cadmium stress treatment, 1 DEG was up-regulated and 5 novel genes were identified. Among all 6 DEGs, DEG-4 was identified as the gene for trans-2,3-enoyl-CoA reductase, DEG-5 was identified as the gene for senescence-associated protein DIN1 and DEG-6 was identified for caffeic acid O-methyltransferase. All the up-regulated genes may play a role in copper and cadmium stress tolerance in alfalfa.

본 연구에서는 알팔파를 이용하여 사일리지 제조시 L. plantarum KCC-10 및 KCC-19를 접종하여 사일리지의 사료가치, 품질 및 미생물상의 변화를 조사하였다. 알팔파 사일리지 제조시 조단백질, ADF, NDF, TDN 함량 그리고 in vitro 건물소화율은 무접종구와 비슷하였다. L. plantarum KCC-10 및 KCC-19 접종구는 무접종구에 비해 알팔파 사일리지의 pH는 감소 되었으나 (p>0.05), 젖산함량은 L. plantarum KCC-10 및 KCC-19 접종구에서 증가하였다(p<0.05). 그러나 초산 및 낙산 함량은 젖산균 접종구에서는 증가하였으나 통계적인 차이는 보이지 않았다 (p>0.05). 알팔파 사일리지 젖산균 단독 처리구의 젖산균 수는 무접종구에 비해 현저하게 증가하였다 (p<0.05). 이상의 결과를 요약해 보면 알팔파 사일리지 제조시 L. plantarum KCC-10 및 KCC-19 접종에 의해 사일리지의 젖산함량과 젖산균수가 증가되었다.

In order to reveal the aluminum (Al) stress tolerance mechanisms in alfalfa plant at low pH soil, a proteomic approach has been conducted. Alfalfa plants were exposed to Al stress for 5 days. The plant growth and total chlorophyll content are greatly affected by Al stress. The malondialdehyde (MDA) and H2O2 contents were increased in a low amount but free proline and soluble sugar contents, and the DPPH-radical scavenging activity were highly increased. These results indicate that antioxidant activity (DPPH activity) and osmoprotectants (proline and sugar) may involve in ROS (H2O2) homeostasis under Al stress. In proteomic analysis, over 500 protein spots were detected by 2-dimentional gel electrophoresis analysis. Total 17 Al stress-induced proteins were identified, of which 8 protein spots were up-regulated and 9 were down-regulated. The differential expression patterns of protein spots were selected and analyzed by the peptide mass fingerprinting (PMF) using MALDI-TOF MS analysis. Three protein spots corresponding to Rubisco were significantly down-regulated whereas peroxiredoxin and glutamine synthetase were up-regulated in response to Al stress. The different regulation patterns of identified proteins were involved in energy metabolism and antioxidant / ROS detoxification during Al stress in alfalfa. Taken together, these results provide new insight to understand the molecular mechanisms of alfalfa plant in terms of Al stress tolerance.

Alfalfa (Medicago sativa L.) is one of the most important forage legumes in the world. It has been demanded to establish the efficient transformation system in commercial varieties of alfalfa for forage molecular breeding and production of varieties possessing new characteristics. To approach this, genetic transformation techniques have been developed and modified. This work was performed to establish conditions for effective transformation of commercial alfalfa cultivars, Xinjiang Daye, ABT405, Vernal, Wintergreen and Alfagraze. GUS gene was used as a transgene and cotyledon and hypocotyl as a source of explants. Transformation efficiencies differed from 0 to 7.9% among alfalfa cultivars. Highest transformation efficiencies were observed in the cultivar Xinjiang Daye. The integration and expression of the transgenes in the transformed alfalfa plants was confirmed by polymerase chain reaction (PCR) and histochemical GUS assay. These data demonstrate highly efficient Agrobacterium transformation of diverse alfalfa cultivars Xinjiang Daye, which enables routine production of transgenic alfalfa plants.

The present study was planned to analyze the nutritional quality, microbial counts and fermentative acids in Italian ryegrass (IRG) 80% and alfalfa 20% (IRG-HV) mediated silage inoculated with lactic acid bacteria (LAB) as a probiotic strain for 3 months. Crude protein (CP), acid detergent fiber (ADF), and neutral detergent fiber (NDF), total digestible nutrient (TDN) and In-vitro dry matter digestibility (IVDMD), lactic acid bacteria (LAB), yeast and fungi counts and fermentation metabolites such as lactic acid, acetic acid and butyric acids were analyzed. The result shows that the nutritional quality and metabolite profiles of silage were significantly improved with LAB. For microbial counts, LAB showed dominant followed by yeast as compared with control silage. The pH of the silage also reduced significantly when silage inoculated with LAB. The result confirmed that silage preparation using different crops with L. plantarum inoculation is most beneficial for the farmers.

본 연구에서는 IRG-Alfalfa 사일리지 제조시 젖산균과 클로렐라를 처리하여 사일리지의 사료가치, 품질 및 미생물상의 변화를 조사하였다. IRG-Alfalfa 사일리지 제조시 대조구와 클로렐라와 젖산균 공용 처리구의 조단백질 함량은 비슷한 수준을 보였으며 섬유소와 TDN 함량 그리고 in vitro 건물소화율 처리구에서는 차이를 나타나지 않았다. IRG-Alfalfa 사일리지 제조시 클로렐라와 젖산균 공용 처리시 pH가 감소되었다. 젖산함량은 대조구에 비해 젖산균 단독처리구에서 증가되었으며(p<0.05), 클로렐라와 젖산균 공용 처리구에서는 젖산함량이 현저하게 증가하였다(p<0.05). 그리고 젖산균 단독처리구의 젖산균 수는 대조구에 비해 현저하게 증가하였으며(p<0.05). 또한 클로렐라와 젖산균 공용처리구도 현저하게 증가하였다(p<0.05).이상의 결과를 요약해 보면 IRG-Alfalfa 사일리지 제조시 젖산균과 클로렐라 처리에 의해 사일리지의 품질이 향상되었다.

자운영 포장에서 알팔파바구미 성충의 발생양상과 1세대 성충의 밭작물 유식물에 대한 기주선호성을 조사하였다. 자운영 포장에서 포충망에 의한 알팔파바구미의 월동 성충은 3월 하순부터 발생하기 시작하여 4월 중순에 피크를 나타내었으며, 1세대 성충은 4월 하순부터 발생하기 시작하여 5월 하순에 피크를 나타내었다. 하지만 월동 성충의 발생량은 매우 적었고, 1세대 성충의 발생량은 매우 많았으나, 연차간 변이가 심하였다. 온실에서 배추 및 콩 등 11종의 유식물에 대한 알팔파바구미 1세대 성충의 7일 차 발생충수는 배추>콩>케일 순으로 유의하게 많았으며, 조, 땅콩, 기장, 가지 및 상추에서 매우 적었고, 옥수수, 쑥갓, 수수에서는 전혀 발생하지 않았다. 7일 차 유식물의 피해주율도 배추(60.0%)>콩(50.0%)>케일(30.0%)>조(16.7%), 기장(13.3%)>상추(3.3%) 순으로 유의하게 심하였다. 한편, 자운영포장에서 11종의 유식물에 대한 7일 차 피해주율은 배추(46.7%), 콩(43.3%)>케일(23.3%)>조(13.3%), 기장(13.3%) 순으로 유의하게 심하여 온실에서의 결과와 같은 경향을 나타내었다. 따라서 알팔파바구미의 기주선호성은 녹비작물포장 주변에서 밭작물의 안전한 재배를 위한 작물선택의 참고자료로 활용될 수 있을 것이다.

 ,  , This study was conducted to evaluate the scouting methods for larva and adults of the alfalfa weevil, Hypera postica Gyllenhal on Chinese milkvetch, Astragalus sinicus L. in the field. Three sampling methods, shake-bucket, shake-picking, and a sweeping net were evaluated for collecting alfalfa weevil larvae. We found significant differences among scouting methods and date in all survey fields. Sweep-net sampling was less able to detect small larvae than large larvae, which were not detected until early April whereas the shake-bucket, and shake-picking methods efficiently collected larvae from middle March. A Pitfall trap with three different baits - no bait, kidney bean seeds and sprouting kidney beans were compared for collection efficiency of alfalfa weevil adults. Collection efficiencies were significantly different by bait(p<, 0.018). Traps baited with sprouting kidney beans were the most efficient for collecting the alfalfa weevil. The number of alfalfa weevil caught was not significantly different between kidney bean seed and no bait in the pitfall trap. Therefore, the shake-bucket method for larvae and a pitfall trap baited with sprouting kidney beans for adults are recommended for scouting of the alfalfa weevil in a Chinese milkvetch field.

Alfalfa weevil, Hypera postica Gyllenhal, is regarded as one of the most important insect pests on Chinese milk vetch and hairy vetch. The insect usually feed on leaves, stems and pods of the green manure which causes severe reduction of biomass. This study was conducted to observe the occurrence of H. postica larva and adult on Chinese milk vetch in 2009 to 2010, and to evaluate host plants preference of 11 different seedlings in 2010, Milyang, Korea. Occurrence of H. postica larvae on Chinese milk vetch was observed to increase from early April clearly and its peak population was showed during middle to late April. The 1st generation of newly emerged adult of H. postica was appeared on 20th May. Newly emerged H. postica adult was fed on green manure such as Chinese milk vetch and hairy vetch as well as various seedlings located at nearby upland. And then the insects were migrated into aestivating places in middle to late of June. Thus, host plant preference of H. postica adults were evaluated with 11 different seedlings using transparent acrylcage and at Chinese milk vetch field. As a result, H. postica adult preferred on Chinese cabbage, followed by soybean, kale, foxtail millet and proso millet. The damaged rating of seedlings were shown to have the same trend with adult occurrence. In particular, few H. postica adults were occurred on eggplant, lettuce, corn, crown daisy and peanut, and thereby those crops were regarded as non-preference plants due to the absence of damage. These informations can be used by upland farmers who sowing from late May to early June for effective management of H. postica.

Red clover (Trifolium pratense L.) and alfalfa (Medicago sativa L.) herbage were harvested, ensiled into laboratory silos and opened after 1,2,3,7,10,15 and 35 d of storage. The pH and nitrogen distribution were detected in all ensiling stages. The results showed that red clover silage have significant lower pH than alfalfa silage at every stage of ensiling. The non-protein N (NPN) concentration of these two forages kept increasing along with the ensiling processing and red clover silage has significant lower accumulation of NPN than alfalfa throughout the whole fermentation. Similar trends were displayed in the concentrations of ammonia N (NH₃-N) and free amino acid N (FAA-N). Peptide-N exhibited rapid increase at the first few days, followed by decline and then increased again.