Cadmium (Cd) toxicity is a serious limitation for agricultural production. In this study, we explored tolerance mechanism associated with Cd toxicity tolerance in alfalfa plants. We used three distinct alfalfa cultivars M. sativa cv. Vernal, M. sativa cv. Zhung Mu, and M. sativa cv. Xing Jiang Daye in this study. Cd showed declined chlorophyll score in Xing Jiang Daye compared with Zhung Mu and Vernal. No significant change observed among the cultivars for root and shoot length. Atomic absorption spectroscopy analysis demonstrated a significant accumulation of Cd, Fe, S and PC in distinct alfalfa cultivars. However, Zhung Mu and Xing Jiang Daye declined Cd accumulation in root, where Fe, S and PC incremented only in Zhung Mu. It suggests that excess Cd in Zhung Mu possibly inhibited in root by the increased accumulation of Fe, S and PC. This was further confirmed by the response of Fe (MsIRT1) and S transporters (MsSULTR1;2 and MsSULTR1;3), and MsPCS1 genes associated with Fe, S and PC availability and translocation in roots and shoots. It suggests that specially the transcript signal inducing the responses to adjust Cd especially in Zhung Mu. This finding provides the essential background for further molecular breeding program for forage crops.
Soil acidity or alkalinity are serious limitations for crop production. The purpose of this study was to clarify the negative effects of extreme pH stress (low and high) on alfalfa vegetative growth (VG) and biomass accumulation (BA). Two-week-old alfalfa seedlings were exposed to different pH (4.0, 4.5, 7.0, 8.0 and 8.5, respectively) levels for 72 hours. Alfalfa grown at pH 4.0 and 8.5 significantly reduced VG and BA, wherein as neutral pH (7.0) comparably exhibited better growth and biomass yield. These results indicate that extreme acidic or alkaline level are critical limiting factors for growth and biomass yield in alfalfa.
Tall fescue (Festuca arundinacea Schreb.) is an important cool season forage plant that is not well suited to extreme heat, salts, or heavy metals. To develop transgenic tall fescue plants with enhanced tolerance to abiotic stress, we introduced an alfalfa Hsp23 gene expression vector construct through Agrobacterium-mediated transformation. Integration and expression of the transgene were confirmed by polymerase chain reaction, northern blot, and western blot analyses. Under normal growth conditions, there was no significant difference in the growth of the transgenic plants and the non-transgenic controls. However, when exposed to various stresses such as salt or arsenic, transgenic plants showed a significantly lower accumulation of hydrogen peroxide and thiobarbituric acid reactive substances than control plants. The reduced accumulation of thiobarbituric acid reactive substances indicates that the transgenic plants possessed a more efficient reactive oxygen species-scavenging system. We speculate that the high levels of MsHsp23 proteins in the transgenic plants protect leaves from oxidative damage through chaperon and antioxidant activities. These results suggest that MsHsp23 confers abiotic stress tolerance in transgenic tall fescue and may be useful in developing stress tolerance in other crops. Keywords: Tall Fescue; Forage; Agrobacterium tumefaciens; MsHsp23; Transformation
Salt stress is one of the most limiting factors that reduce plant growth, development and yield. However, identification of salt-inducible genes is an initial step for understanding the adaptive response of plants to salt stress. In this study, we used an annealing control primer (ACP) based GeneFishing technique to identify differentially expressed genes (DEGs) in Italian ryegrass seedlings under salt stress. Ten-day-old seedlings were exposed to 100 mM NaCl for 6 h. Using 60 ACPs, a total 8 up-regulated genes were identified and sequenced. We identified several promising genes encoding alpha-glactosidase b, light harvesting chlorophyll a/b binding protein, metallothionein-like protein 3B-like, translation factor SUI, translation initiation factor eIF1, glyceraldehyde-3-phosphate dehydrogenase 2 and elongation factor 1-alpha. These genes were mostly involved in plant development, signaling, ROS detoxification and salt acclimation. However, this study provides new molecular information of several genes to understand the salt stress response. These genes would be useful for the enhancement of salt stress tolerance in plants.
국내에서 육성한 톨 페스큐 신품종 푸르미의 종자증식 기술개발과 종자생산성 및 채종짚의 사료가치 평가를 통해 종자생산 기술체계를 확립하고자 국립축산과학원 초지사료과 시험포장에서 파종시기별 종자생산성을 조사하였다. 파종시기에 따른 출수기의 차이는 약 2~3일정도 차이를 보였으며 종자수량의 결정에 중요한 요소인 단위면적당 이삭수는 9월 17일 파종이 692본/$m^2$로서 가장 많았고 이보다 10일 빠르거나 늦은 경우 점차 감소하는 경향을 보였다. 채종 종자의 천립중도 2 g 내외로 발아율도 80~83%로 처리구간의 큰 차이는 나타나지 않았다. 채종 종자수량의 수량성은 파종시기가 9월 17일 경우 1,711 kg/ha로 가장 많았으며 파종시기가 이보다 10일 빠르거나 늦은 경우 생산된 종자의 수량은 감소하는 경향이 나타났다. 생초 수량과 건물 수량은 파종시기가 9월 17일 파종시 각각 63,556 kg/ha, 16,926 kg/ha로서 가장 많았으며 파종시기가 10일 간격씩 늦어질수록 건물수량도 점차 감소하는 경향을 보였다. 반면 채종짚의 사료가치는 유의적인 차이를 나타내지 않았다. Tall fescue (Festuca arundinacea Schreb.) is one of cool-season adapted perennial grass species and has been cultivated worldwide as a important forage and lawn grass. In this study, we evaluated the seed production of new tall fescue cultivar, 'Purum', which was cultivated in Korea. Agronomic aspects of tall fescue were determined using seed production and forage nutritive value of straw at the National Institute of Animal Science, RDA from 2012 to 2013. The heading date according to the seeding time displayed 2~3 days differences. The harvested seed yields were the largest (1,711 kg/ha) when seeding date at September $17^{th}$, whereas it decreased when planted ten days earlier or later than at that time. The thousand-grain weight of gathering seeds was 2 g, and the germination rate was 80 to 83% showing no significant differences among the different seeding times. The yield of fresh weight and dry matter were the largest (63,556 kg/ha and 16,926 kg/ha, respectively) when planted at September $17^{th}$. The dry matter yield was gradually decreased with delaying seeding time as 10 days period. However, nutritive values of straw did not showed significant differences.
이탈리안 라이그라스의 발아 및 초기 생육단계에서 내염성 수준 정도를 조사하기 위하여 in vitro 조건에서 다양한 염농도 조건에서 그린팜, 플로리다80, 코원어리 및 화산101호 품종을 이용하여 염해에 대한 반응을 발아율과 산화 스트레스 관련 효소들의 활성을 통해 조사하였다. 저농도의 NaCl 처리구에서는 모든 품종에서 발아율의 차이가 1~3% 정도 감소하는 경향을 보였으며 250 mM 이상의 NaCl 처리구에는 화산101호 품종을 제외한 모든 품종에서 50% 이상의 발아율 감소를 보였다. 또한, NaCl 농도가 높아질수록 발아에 걸리는 시간이 지연되었으며, 350 mM NaCl이 첨가된 처리구에는 화산101호 품종만이 치상 10일후에 발아가 시작하여 12% 정도 발아하였다. NaCl 농도에 따른 shoot과 root의 길이와 생초와 건조 후 무게의 변화도 만생종으로 갈수록 더 생육이 우수한 것으로 타나났다. 품종별간의 잎 조직에서 항산화효소 활성을 분석 결과 고염 조건 (150 mM NaCl)에서 POD 활성은 대조군과 유사한 수준으로 확인 되었다. 반면 APX 활성은 플로리다80과 코윈어리 품종에서는 증가하는 경향을, 화산101 품종에서는 감소하는 경향이 나타났으나 유의적인 수준에서 차이는 확인되지 않았다. 반면 그린팜 품종에서는 유의적인 수준에서 고염 처리에 의한 APX 활성이 감소되는 경향이 나타났다. 지속적으로 in vitro screening을 통해 선발된 계통들은 향후 특성평가와 인공교배를 통해 내염성 이탈리안 라이그라스 개발에 필요한 유용한 육종모재로 활용될 수 있을 것으로 판단된다. This study was conducted to evaluate the performance of Italian ryegrass cultivars for salt tolerance under in vitro condition. Italian ryegrass cultivars such as Greenfarm, Florida80, Kowinearly, and Hwasan101 were tested for their tolerance to various sodium chloride levels (0, 50, 150, 250, and 350 mM). The seed germination, growth, and activities of antioxidant enzymes were investigated under salt treatment. Physiological traits such as seed germination percentage, germination period, shoot and root length, and dry weight were suppressed under entire salt stress conditions. The results indicated that the highest germination percentage and shoot and root length were recorded at normal conditions. Increased sodium chloride levels caused a significant reduction in the seed germination and growth rate. Among the four tested cultivars, Italian ryegrass 'Hwasan101' could be considered as salt tolerant owing to its higher germination percentage, better seedling growth and antioxidant activities under salinity stress, whereas Greenfarm cultivar was more sensitive. The selection of Italian ryegrass cultivars for greater tolerance to saline environment would allow greater productivity from large saline lands.
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