苜蓿原生质培养及体细胞杂交技术的应用

阐述了植物原生质体培养和体细胞杂交技术以及苜蓿Medicago spp.体细胞胚胎发生特性,综述了该技术在苜蓿育种研究中的应用状况,同时介绍了我国苜蓿研究领域原生质体培养和体细胞杂交技术的研究进展.     

沙引发对紫花苜蓿种子盐逆境下发芽及幼苗生理生化变化的影响

试验以紫花苜蓿品种维多利亚和金皇后种子为材料，研究了沙引发处理对高盐(0．8％NaCl)逆境下种子发芽及幼苗生理生化变化的影响。结果表明，沙引发处理能显著提高紫花苜蓿两个品种种子的发芽率、发芽势、发芽指数和活力指数，缩短平均发芽时间，显著增加幼苗干重，同时提高了过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性，减少丙二醛(MDA)的积累。说明沙引发提高了紫花苜蓿种子的活力和抗盐胁迫能力，促进了盐逆境下种子的萌发和幼苗生长。     

Influence of P and K fertilization on Phytophthora root rot or excess soil water injury of alfalfa cultivars

Abstract

Phytophthora root rot (PRR) is an important disease of alfalfa (Medicago sativa L.) in wet soils. Excess soil water (ESW) stress is required for the successful infection and development of PRR. The effects of P at 0, 40, and 80 μg.g^‐1 soil and K at 0, 120, and 240 μg.g^‐1 soil (in factorial combinations) on the severity of PRR or ESW stress was investigated on a PRR‐susceptible (Iroquois) and a resistant (Oneida) alfalfa cultivar in a greenhouse trial. P and K applications significantly increased the dry weight of tops prior to imposition of PRR or ESW stress. However, plants subjected to PRR or ESW stress did not respond significantly to P or K and there was no significant effect of P or K on the severity of PRR or ESW injury. The plants subjected to PRR or ESW stress had lower tissue P concentration than those under stress‐free conditions. Increasing P additions resulted in a significant increase in P concentration of Iroquois plants, while in Oneida the response was non‐significant. The response of K concentration in the plant to increasing K additions was dependant on P levels.     

新牧一号苜蓿MvP5CS基因的克隆和功能分析

为研究苜蓿（Medicago sativa）耐盐分子机制，为抗逆分子育种提供依据，通过同源克隆和RT PCR技术克隆了新牧一号杂花苜蓿（M.varia Xinmu 1）的MvP5CS基因， MvP5CS全长2 148 bp，Genbank登录号为：EU371644。荧光定量PCR分析发现，在盐胁迫下MvP5CS基因表达明显上调，说明 MvP5CS与苜蓿的耐盐性相关。利用农杆菌介导的叶盘转化法，将MvP5CS基因成功转入烟草（Nicotiana tabacum），盐胁迫下转MvP5CS基因T1代烟草萌发率和根长均高于非转基因烟草。表明新牧一号苜蓿P5CS基因能够提高转基因烟草的耐盐性。     

EFFECTS OF pH ON GROWTH AND NODULATION OF TWO FORAGE LEGUMES

A study was conducted to compare the responses to acid pH of Medicago sativa and Lotus glaber, two forage legumes with different environmental requirements, either supplied with inorganic nitrogen (N) or inoculated with different strains of their nodule bacteria. Medicago sativa showed, in both treatments, a significant reduction in total dry weight at pH below 6.0. In contrast, Lotus glaber grew equally well at all the pHs assayed in the presence of adequate N. Under inoculated conditions, in the absence of N supply, plant growth was dependent on the bacterial strain used. When the ability of each strain to multiply in culture medium was examined, it was observed that Sinorhizobium meliloti strains showed a pH-sensitive response that inhibited growth at pH 4.0, whereas Mesorhizobium loti strains showed normal growth at this pH. These results suggest that for the effectiveness of Mesorhizobium loti–Lotus glaber symbiosis in acid soils the major factor to be considered is the tolerance of the bacterial strain to acid conditions, while the limiting factors for the Sinorhizobium meliloti–Medicago sativa symbiosis are the sensitivity to low pH of both the plant and its bacteria.     

Effectiveness of Coniothyrium minitans and Trichoderma atroviride in suppression of sclerotinia blossom blight of alfalfa

The effects of the mycoparasites Coniothyrium minitans and Trichoderma atroviride on the suppression of alfalfa blossom blight caused by Sclerotinia sclerotiorum were evaluated under indoor and field conditions. When T. atroviride (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) were applied to detached young alfalfa florets, T. atroviride effectively inhibited saprophytic growth of S. sclerotiorum, whereas C. minitans showed no inhibition under the same conditions. When T. atroviride (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) was applied to young alfalfa petals in vivo just after pollination, the percentage of pod formation was higher for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum, and the percentage of pod rot was lower for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum. However, when they were applied to senescent petals attached to developing pods of alfalfa at 9·2 × 10⁴ conidia/floret together with S. sclerotiorum at 4·5 × 10³ ascospores/floret at 14 days after pollination, C. minitans was more effective than T. atroviride in suppressing sclerotinia pod rot and seed rot of alfalfa. Field experiments showed that three applications of C. minitans (5·4 × 10⁶ conidia mL⁻¹) or T. atroviride (5·4 × 10⁶ conidia mL⁻¹) at a 7-day interval to blossoms of alfalfa effectively suppressed sclerotinia pod rot in two out of three annual trials. Coniothyrium minitans effectively suppressed sclerotinia seed rot in all three years, whereas T. atroviride was not effective against seed rot in any of the trial years. The efficacy of C. minitans was not significantly different (P > 0·05) from benomyl (250 µg ai mL⁻¹). This study suggests that C. minitans has potential as a biocontrol agent to control blossom blight of alfalfa caused by S. sclerotiorum.     

内蒙古中东部野生扁蓿豆种子硬实性的研究

扁蓿豆是抗逆性极强的优良牧草,但其种子发芽率低,具有65 %～75 %的硬实。在恒温25℃时发芽率较高,在变温20 ～30℃时发芽率最高,若温度超过30℃或低于15℃时发芽被抑制,但短时间高温处理有利于促进发芽。采用机械或浓硫酸处理,发芽率分别比对照高40.6 %和45.6 %。野生种子的硬实率与采集地气温,特别是与7月份温度有着很高的正相关关系,而且不同产地的种子表现出较大的差异,以赤峰市北部采到的居群硬实率最高,比呼和浩特市采集的居群高9 %。随着储藏年限的推移,种子的硬实率不断下降,发芽率增加,储藏3.5年的种子发芽率达到了67.0 %,表明扁蓿豆种子采集后通过室温储藏可以达到提高发芽率的效果。     

环峡南苜蓿种子生产技术研究

对影响环峡南苜蓿种子产量的磷肥、钾肥、浇水和地膜条件4个因素进行正交组合试验,结果表明: 磷肥、钾肥、磷钾肥互作对种子产量的影响较大,其次为地膜条件、磷肥浇水互作,浇水的影响最小。试验结果和生产成本综合考虑,最适宜的处理组合为:钙镁磷450 kg/hm2+硫酸钾150 kg/hm2+不铺地膜+不浇水。     

天蓝苜蓿组织培养及再生体系的研究

以野生天蓝苜蓿种子发育5～7d无菌苗的子叶、下胚轴和种子苗叶片为外植体,对其愈伤组织诱导及其分化的基本培养与外源激素组合和浓度配比进行试验。结果表明:对子叶、下胚轴和叶片愈伤组织诱导最适宜的基础培养基分别是MS、B5和SH;激素的种类及其浓度配比对于愈伤组织的诱导因不同的外植体有很大的差异。其中,子叶、下胚轴以MS+NAA(0.5～1.0 mg/L)+6-BA(0.7mg/L)效果较好,叶片以SH+NAA(0.5～1.0mg/L)+2,4-D(1.5～5 mg/L)+6-BA(0.7mg/L)较好,少量下胚轴在MS+NAA(0.5mg/L)+6-BA(0.7mg/L)和MS+NAA(0.5mg/L)+KT(0.5 mg/L)的培养基中直接分化出胚状体,并成功发育成植株。在培养基Whb5上,子叶、下胚轴和叶片的分化都得到了很好的效果,分化率分别为43%、57%、40%,15～30d后都能再生植株。     

扁蓿豆生长发育规律的研究

通过4年的种植观察发现,种植第1年扁蓿豆和苜蓿生长都较缓慢,扁蓿豆的越冬率在l～4年内随着生育年限的增加而提高,而苜蓿的越冬率比扁蓿豆稍低.返青都在4月末,从返青到成熟期的生育天数扁蓿豆为120d,苜蓿为103d.生活第2年的扁蓿豆草层最大鲜量出现在30～40cm处,其中叶量占52.4%;生活第2年和生活第3年的苜蓿草层结构最大鲜量都出现在40～50cm处,分别为50.6g和55.2g,比扁蓿豆分别少了12.4g和7.2g.种子产量第3年最高,扁蓿豆为188.4kg/hm2,苜蓿为211.95kg/hm2.扁蓿豆的生育期较长,第1年为150d,第2年为125d,第3年为120d;而生活第2年的苜蓿生育期为120d,第3、4年都为103d.