The Changes in Anti-oxidant Activity of Roots in Wintering Period under Single Sowing of Alfalfa and Mixed Sowing of Alfalfa and Bromus inermis Leyss

This paper aimed to study the change in MDA content and antioxidant enzyme activity of roots in wintering period under single sowing of alfalfa and mixed sowing of alfalfa and Bromus inermis Leyss.By the single sowing of 4 alfalfa varieties with different cold resistance and mixed sowing of alfalfa varieties and Bromus inermis Leyss,the changes in MDA content,SOD,POD and CAT activity of alfalfa roots throughout the wintering period were measured.Results indicated that after single sowing of alfalfa and mixed sowing of alfalfa and Bromus inermis Leyss,the MDA content of roots showed an up-down-up trend with temperature; the CAT activity increased with the decreasing temperature but decreased when the temperature rose in the spring of the following year; the SOD and POD activity showed an increasing trend with sharp decline in temperature,and decreased when the temperature continued to decline,but increased with alfalfa reviving in the following year.The enzymic activity of the same variety under mixed sowing was higher than under single sowing.The membership function was used for comprehensive evaluation of cold resistance,and the cold resistance under different treatments was in the order of Wega7F + Bromus inermis Leyss >Wega7F > Xunlu + Bromus inermis Leyss > Xunlu > Aohan + Bromus inermis Leyss > Aohan > WL319HQ + Bromus inermis Leyss > WL319HQ.This was of great significance to the study on cold resistant alfalfa breeding and cultivation in cold areas of northern China.

Responses of Caryopsis Germination, Seedling Emergence, and Development to Sand Water Content of Agropyron cristatum （L.） Gaertn. and Bromus inermis Leyss.

Responses of caryopsis germination, seedling emergence, and development of Agropyron cristatum （L.） Gaertn. （Gramineae） and Bromus inermis Leyss. （Gramineae）, two dominant perennial grasses in the Otindag Sandland of China, to different sand water content （SWC; 1%, 2%, 3%, 4%, 6%, 8%, 12%, 16%, and 20%） were studied comparatively. The results showed that the germination responses of the two grasses to SWC were similar （i.e. caryopses could not germinate when the SWC was below 3%; at SWC ranging from 3% to 12%, the higher the SWC, the higher the germination percentage; and at a SWC of 12%-20%, germination reached similarly high percentages）. At a sand burial depth of 0.5 cm, the threshold of SWC for seedling emergence was 6% forA. cristatum and 8% forB. inermis; at 12%-20% SWC, the seedling emergence of both species reached similarly high percentages. The seedling growth responses of these two species to SWC gradients were different. For A. cristatum, the biomass of seedlings increased with SWC from 6% to 12%, and decreased with SWC from 12% to 20%. For B. inermis, the biomass of seedlings always increased with SWC from 8% to 20%. The results also showed that the seedlings of both species allocated more biomass to the roots with decreases in SWC. The SWC changes from April to October in natural microhabitats of both species suggested that the SWC may play an important role in caryopsis germination, seedling emergence, and the growth characteristics of the two grasses. The responses of caryopsis germination, seedling emergence, and the growth characteristics of these two species to SWC may determine their distribution pattems in the Otindag Sandland.

Effect of nitrogen fertilization on yield, N content, and nitrogen fixation of alfalfa and smooth bromegrass grown alone or in mixture in greenhouse pots

Planting grass and legume mixtures on improved grasslands has the potential advantage of realizing both higher yields and lower environmental pollution by optimizing the balance between applied N fertilizer and the natural process of legume biological nitrogen fixation. However, the optimal level of N fertilization for grass-legume mixtures, to obtain the highest yield, quality, and contribution of N2 fixation, varies with species. A greenhouse pot experiment was conducted to study the temporal dynamics of N2 fixation of alfalfa （Medicago sativa L.） grown alone and in mixture with smooth bromegrass （Bromus inermis Leyss.） in response to the addition of fertilizer N. Three levels of N （0, 75, and 150 kg ha-1） were examined using 15N-labeled urea to evaluate N2 fixation via the 15N isotope dilution method. Treatments were designated NO （0.001 g per pot）, N75 （1.07 g per pot） and N150 （2.14 g per pot）. Alfalfa grown alone did not benefit from the addition of fertilizer N; dry matter was not significantly increased. In contrast, dry weight and N content of smooth bromegrass grown alone was increased significantly by N application. When grown as a mixture, smooth bromegrass biomass was increased significantly by N application, resulted in a decrease in alfalfa biomass. In addition, individual alfalfa plant dry weight （shoots＋roots） was significantly lower in the mixture than when grown alone at all N levels. Smooth bromegrass shoot and root dry weight were significantly higher when grown with alfalfa than when grown alone, regardless of N application level. When grown alone, alfalfa＇s N2 fixation was reduced with N fertilization （R2=0.9376,P=0.0057）. When grown in a mixture with smooth bromegrass, with 75 kg ha-1 of N fertilizer, the percentage of atmospheric N2 fixation contribution to total N in alfalfa （%Ndfa） had a maximum of 84.07 and 83.05% in the 2nd and 3rd harvests, respectively. Total 3-harvest %Ndfa was higher when alfalfa was grown in a mixture than when grown alone （shoots： ｜t｜=3.39, P=0.0096; root： ｜t｜=3.57, P=0.0073）. We believe this was due to smooth bromegrass being better able to absorb available soil N （due to its fibrous root system）, resulting inlower soil N availability and allowing alfalfa to develop an effective N2 fixing symbiosis prior to the 1st harvest. Once soil N levels were depleted, alfalfa was able to fix N2, resulting in the majority of its tissue N being derived from biological nitrogen fixation （BNF） in the 2nd and 3rd harvests. When grown in a mixture, with added N, alfalfa established an effective symbiosis earlier than when grown alone; in monoculture BNF did not contribute a significant portion of plant N in the N75 and N150 treatments, whereas in the mixture, BNF contributed 17.90 and 16.28% for these treatments respectively. Alfalfa has a higher BNF efficiency when grown in a mixture, initiating BNF earlier, and having higher N2 fixation due to less inhibition by soil-available N. For the greatest N-use-efficiency and sustainable production, grass-legume mixtures are recommended for imDrovino orasslands, usino a moderate amount of N fertilizer （75 kq N ha-l） to provide optimum benefits.

Elevation difference of Ca^(2+) levels in young leaf cells of bromegrass and induced cold-tolerant enhancement under different controlled chilling temperatures

The changes of Ca2＋ levels in young leaf cells of bromegrass under different controlled chilling temperatures were inves- tigated by an antimonite precipitation cytochemical method. The main results were as follows： under 25/20℃ （day/night） tempera- ture and 14 h photoperiod, electron-dense Ca2＋ antimonite precipitates, indicators of Ca2＋ localization, were mainly localized in the vacuoles, cell walls and intercellular spaces; few Ca2＋ deposits were observed in the cytosol and nuclei. After a 3℃ chilling treatment for 3 h, many Ca2＋ precipitates appeared in the cytosol and nuclei, indicating that Ca2＋ influx had occurred in the cytosol and nuclei. When the 3℃ treatment was prolonged to 8 h, more Ca2＋ deposits appeared in the nuclei and cytosol, but the amount of Ca2＋ deposits in both the cytosol and nuclei decreased markedly after a 24 h treatment and most Ca2＋ deposits were returned to the vacuoles and intercellular spaces after an 8 d treatment. When bromegrass was exposed to 7℃ for 3 h, the Ca2＋ distribution in the cells had no visible changes, compared with that of the 25/20℃ grown control plants. However, when the chilling treatment of 7℃ was prolonged to 8 h, a Ca2＋ influx occurred, where many Ca2＋ deposits were observed in the nuclei and cytosol. More Ca2＋ deposits appeared in the nuclei and cytosol after a 24 h treatment, but the amount of Ca2＋ deposits in the cytosol and nuclei was reduced markedly after an 8 d treatment. After a 14 d treatment, the remaining low level of Ca2＋ was recovered in both the cytosol and nuclei and the Ca2＋ deposits were again located in the vacuoles and the intercellular spaces. The dynamics of subcellular Ca2＋ localization in young leaf cells of bromegrass during a 12℃ chilling treatment were similar to those of the 7℃ treatment. Besides, the results showed that the frost tolerance ofbromegrass exposed to 3℃ for 8 d increased by 6℃, for 7℃ and 8 d by 4℃ and for 12℃ and 14 d by 3℃, compared with the controls. Finally, the relationship between different Ca2＋ dynamics and induced frost tolerance was also explored.

基于ISSR标记的无芒雀麦种质资源遗传多样性

本研究以29份来自国内外无芒雀麦种质材料为研究对象,利用ISSR分子标记进行遗传多样性研究,以期为无芒雀麦种质资源的收集、利用和育种提供了理论依据。试验筛选出16对引物进行PCR扩增,共扩增出273个ISSR条带,其中,多态性条带253个,多态性位点百分率达92.67%,Nei’s基因多样性指数(H)的范围在0.255~0.355之间,平均值为0.310。遗传相似系数范围在0.518~0.902之间,平均值为0.649,说明不同无芒雀麦间存在较大差异。在阈值为0.644处,29份材料被分为4类,UPGMA聚类结果显示,各供试材料间的聚类与其地理来源有一定的相关性,主成分分析与聚类结果一致。ISSR分子标记是遗传多样性研究的有效手段。