紫花苜蓿的花蜜量和访花蜜蜂数量对种子产量的影响

对10个紫花苜蓿品种的花萼直径、花蜜量、访花蜜蜂数量和种子产量进行了研究,结果表明,紫花苜蓿的单位面积花蜜量与访花蜜蜂数量呈极显著正相关(r=0.93**),访花蜜蜂数量与种子产量呈显著正相关(r=0.87*);紫花苜蓿的花蜜量与花萼直径呈极显著正相关(r=0.99**);紫花苜蓿品种赛特(Sitel)和德宝(Derby)在兰州地区能获得高产、优质的种子。     

不同播期对紫花苜蓿生长性状及越冬性的影响研究

采用随机试验设计方法,在宁南旱作农业区进行了不同播期对紫花苜蓿出苗率、生长发育、越冬率和产草量影响的研究。结果表明,在宁南地区人工种植紫花苜蓿的出苗率与播期温度存在极显著正相关关系,而不同播期土壤耕层含水量变化率较小,且基本能满足苜蓿萌发与生长的需要;播期越早,根颈越粗,入土越深,但4月份前播种,出苗率低,群体小且易形成“小老苗”;7月份以后播种。由于生长期短,植株根颈细嫩,越冬率很低或完全冻死。综合不同播期出苗率、产草量和越冬率等因素,认为在宁南地区紫花苜蓿的适宜播种期应在4月30日～6月30日。     

Hg~(2+)对苜蓿叶片的毒害效应

研究了Hg2 + 对苜蓿叶片的毒害效应。结果表明 :随着Hg2 + 浓度的增加和处理时间的延长 ,叶绿素含量下降 ,电导度上升 ;低Hg2 + 浓度及短时间处理 ,超氧阴离子自由基 (O·2 )和丙二醛 (MDA)含量增加 ,SOD、POD和CAT等保护酶活性升高 ,表明膜系统受到了伤害 ;高Hg2 +浓度及长时间处理 ,O·2 和MDA含量下降 ,SOD、POD和CAT等保护酶活性降低 ,表明细胞结构和功能受到了不可逆的伤害。     

杂花和紫花苜蓿原生质体分离培养条件的筛选

以杂花苜蓿‘甘农1号’和紫花苜蓿‘甘农4号’、‘阿尔冈金’及‘清水’4个适宜西北内陆黄土高原地区栽培的苜蓿愈伤组织为材料,研究酶解时间、酶液组合、酶液渗透压、愈伤组织继代培养时间、预处理措施及不同培养方法等对原生质体分离和培养效果的影响,并对培养条件进行优化。结果表明:(1)适宜4个苜蓿品种愈伤组织酶解的最佳预处理措施为0.55mol/L蔗糖或CPW溶液中预质壁分离1h,最佳继代时间均为12d。(2)‘甘农1号’、‘甘农4号’和‘清水’的最佳酶液组合均为2%纤维素酶+0.5%果胶酶+0.3%崩溃酶;‘阿尔冈金’的最佳酶液组合为2%纤维素酶+0.5%果胶酶+0.3%半纤维素酶+0.3%离析酶+0.3%崩溃酶;‘甘农1号’和‘阿尔冈金’的最佳酶解时间为12h,‘甘农4号’和‘清水’分别为14h和10h。(3)适宜4个品种酶解的甘露醇浓度分别为‘甘农1号’0.75mol/L,‘甘农4号’0.65mol/L,‘阿尔冈金’0.6mol/L,‘清水’0.55~0.6mol/L。(4)经液体浅层培养和固液培养方式均可获得4个苜蓿品种的再生愈伤组织,且固液培养法较液体浅层培养法更有利于苜蓿原生质体早期的培养和再生。     

Effects of tissue culture on a highly repetitive DNA sequence (E180 satellite) in Medicago sativa

Genomic DNA extracted from leaves of sixteen alfalfa genotypes, six-week-old callus cultures derived from them and from a suspension culture was used in Southern blots and slot blots probed with alfalfa E180 satellite DNA. The restriction patterns revealed by the E180 probe did not differ among alfalfa genotypes and no differences between the leaf and callus restriction patterns were seen. However, the number of copies of the E180 satellite was lower in the callus samples than in the corresponding leaf samples and significant differences in copy number among callus samples were detected. Genomic stress induced by tissue culture may have caused the reduction in copy number. This phenomenon may be important in the generation of somaclonal variation in alfalfa.     

Down-regulated expression of plant-specific glycoepitopes in alfalfa

Compared with other plant expression systems used for pharmaceutical protein production, alfalfa offers the advantage of very homogeneous N -glycosylation. Therefore, this plant was selected for further attempts at glycoengineering. Two main approaches were developed in order to humanize N -glycosylation in alfalfa. The first was a knock-down of two plant-specific N -glycan maturation enzymes, β1,2-xylosyltransferase and α1,3-fucosyltransferases, using sense, antisense and RNA interference strategies. In a second approach, with the ultimate goal of rebuilding the whole human sialylation pathway, human β1,4-galactosyltransferase was expressed in alfalfa in a native form or in fusion with a targeting domain from N -acetylglucosaminyltransferase I, a glycosyltransferase located in the early Golgi apparatus in Nicotiana tabacum . Both knock-down and knock-in strategies strongly, but not completely, inhibited the biosynthesis of α1,3-fucose- and β1,2-xylose-containing glycoepitopes in transgenic alfalfa. However, recombinant human β1,4-galactosyltransferase activity in transgenic alfalfa completely prevented the accumulation of the Lewis a glycoepitope on complex N -glycans.     

<Emphasis Type="Italic">In vitro</Emphasis> direct organogenesis and regeneration of <Emphasis Type="Italic">Medicago sativa</Emphasis>

A rapid and efficient plant regeneration protocol for a wide range of alfalfa genotypes was developed via direct organogenesis. Through a successive excision of the newly developed apical and axillary shoots, a lot of adventitious buds were directly induced from the cotyledonary nodes when hypocotyl of explants were vertically inserted into modified Murashige and Skoog (MS) medium supplemented with 0.025 mg dm⁻³ thidiazuron (TDZ) and 3 mg dm⁻³ AgNO₃. When the lower part of shoots excised from explants were immersed into the liquid medium with 1.0 mg dm⁻³ α-naphthaleneacetic acid (NAA) for 2 min, and then transferred to hormone free half-strength MS medium, over 83.3 % of the shoots developed roots, and all plantlets could acclimatize and establish in soil. The protocol has been successfully applied to eight genotypes, with regeneration frequencies ranging from 63.8 to 82.5 %.     

Growth of Salmonella on sprouting alfalfa seeds as affected by the inoculum size, native microbial load and Pseudomonas fluorescens 2-79

Aims: To investigate the growth of salmonellae on sprouting alfalfa seeds as affected by the inoculum size, microbial load and Pseudomonas fluorescens 2–79. Methods and Results: Alfalfa seeds pre‐inoculated with ≤10¹–10³ CFU g^?1 of salmonellae and with or without Ps. fluorescens 2–79 were sprouted in glass jars and the population of salmonellae were determined daily for up to 6 days. The population of salmonellae on germinating seeds reached the maximum 2–3 days after sprouting when total bacterial count reached the maximum (10⁹ CFU g^?1). The population of salmonellae on sprouting seeds not treated with Ps. fluorescens 2–79 showed a net increase of 3–4 log units. However, the population of salmonellae on alfalfa seeds treated with Ps. fluorescens 2–79 showed a net increase of only 1–2 log units. Disinfection of seeds with calcium hypochlorite enhanced the growth of salmonellae. Conclusions: Treatment of seeds with Ps. fluorescens 2–79 reduced the growth of salmonellae by 2–3 log units. Significance and Impact of the Study: The potential of Ps. fluorescens 2–79 as a biological agent for use in control of salmonellae on sprouting seeds was demonstrated and warrants further investigation.