Embryo culture of Medicago scutellata and M. sativa

Resistance to the alfalfa weevil (Hypera postica (Gyllenhal)) and the potato leafhopper (Empoasco fabae (Harris)) is lacking in cultivated alfalfa. However, a closely related annual Medicago, Medicago scutellata, possesses dense glandular stem and leaf hairs which provides a mechanism for resistance. Several attempts have been made at transfering the glandular haired traint from M. scutellata to perennial alfalfa with limited success. Earlier studies have shown that one reason for the lack of success is embryo abortion. Therefore, this study was initiated to observe zygotic embryo-genesis and to develop an embryo rescue technique for M. scutellata and M. sativa. Observations of zygotic embryogenesis showed that the two species are similar in morphology and can be described from youngest to oldest as globular, heart, torpedo, and hook shaped embryos. M. sativa embryos are smaller than M. scutellata embryos and develop three to four days later. Self pollinated M. scutellata (PI 307446) and sib mated M. sativa (Saranac AR) embryos were cultivated on Murashige and (2,4-D), indolacetic acid (IAA), 6-benzylaminopurine (BAP), and kinetic (KIN). Embryos from both species were also cultured on Schenk and Hildebrandt's (SH) basal medium with the addition of L-glutamine and L-proline. The experimental design was a completely randomized factorial for each experiment. Heart and torpedo shaped embryos from M. scutellata grew best (27.5% plantlet recovery) when cultured on MS medium with 0.05 mgl^-1 of both IAA and BAP. After 15 to 30 days on this medium, the embryos had only developed shoots. Therefore, it was necessary to transfer the shoots to MS basal medium without phytohormones for rooting. Rooting occurred in 15 to 30 days and the plantlets could be acclimatized to soil within 2 to 4 weeks. M. sativa embryos grew best (31% plantlet recovery) on SH medium with 50 mM L-glutamine. M. sativa embryos developed both shoots and roots on this medium. This information may now be applied to the development of an embryo culture method for recovering insect resistant hybrids between M. scutellata and M. sativa. Disclaimer statement: Mention of a trademark or proprietary product does not constitute a guarantee or warranty of the product by the USDA, and does not imply its approval to the exclusion of other products that may also be suitable.     

Mass Culturing of Ditylenchus dipsaci to Yield Large Quantities of Inoculum

Methods are described for rearing large quantities of Ditylenchus dipsaci on alfalfa tissues. Nematodes and alfalfa seed were disinfected and nematodes were reared in quantities sufficient to provide a continuous supply of inoculum for our alfalfa-breeding program. Nematodes reproduced best in darkness at 20-25 C. Cultures reached maximum numbers in 3-6 wk.     

Host-parasite Interactions of Pratylenchus scribneri on Selected Crop Plants

Greenhouse tests were conducted to determine the effects of soil temperature and texture on development of Pratylenchus scribneri and the pathogenicity and reproductive rates of this nematode on selected crop plants. In a sandy loam soil, greatest numbers of P. scribneri were found at 30 and 35 C on sudangrass and sugarbeet, respectively. In a silty clay loam, the nematode reproduced best at 35 C on sugarbeet. Higher populations of P. scribneri were found in the sandy loam than silty clay loam soil at corresponding temperatures. In a pathogenicity test, top and root growth of sudangrass and barley were suppressed by the nematode, whereas no significant growth inhibition was found on wheat and alfalfa. Tests with other vegetable and field crops indicated wide variance in nematode reproduction.     

New aspects of grassland recovery in old‐fields revealed by trait‐based analyses of perennial‐crop‐mediated succession

Classical old‐field succession studies focused on vegetation changes after the abandonment of annual croplands or on succession after the elimination of cultivated crops. Perennial‐crop‐mediated succession, where fields are initially covered by perennial crops, reveals alternative aspects of old‐field succession theories. We tested the validity of classical theories of old‐field succession for perennial‐crop‐mediated succession. We formulated the following hypotheses: (1) functional diversity increases with increasing field age; (2) resource acquisition versus conservation trade‐off shifts toward conservation at community level during the succession; (3) the importance of spatial and temporal seed dispersal decreases during the succession; and (4) competitiveness and stress‐tolerance increases and ruderality decreases at community level during the succession. We studied functional diversity, trait distributions and plant strategies in differently aged old‐fields using chronosequence method. We found increasing functional richness and functional divergence, but also unchanged or decreasing functional evenness. We detected a shift from resource acquisition to resource conservation strategy of communities during the succession. The role of spatial and temporal seed dispersal was found to be important not only at the initial but also at latter successional stages. We found an increasing stress‐tolerance and a decreasing ruderality during succession, while the competitiveness remained unchanged at the community level. Despite the markedly different starting conditions, we found that classical and perennial‐crop‐mediated old‐field successions have some similarities regarding the changes of functional diversity, resource acquisition versus conservation trade‐off, and seed dispersal strategies. However, we revealed also the subsequent differences. The competitive character of communities remained stable during the succession; hence, the initial stages of perennial‐crop‐mediated succession can be similar to the middle stages of classical old‐field succession. Moreover, the occupied functional niche space and differentiation were larger in the older stages, but resources were not effectively utilized within this space, suggesting that the stabilization of the vegetation requires more time.     

苹果炭疽叶枯病菌致病力分析及苹果种质抗病性鉴定

苹果炭疽叶枯病是由胶孢炭疽菌(Colletotrichum gloeosporioides)引起的一种真菌病害,现已上升为世界苹果生产中的主要病害之一。了解不同来源的苹果炭疽叶枯病菌致病力差异及明确苹果种质资源对苹果炭疽叶枯病的抗性,对品种选育、品种合理布局以及控制病害的流行具有重要的参考价值。本研究对不同来源的79株病原菌进行了室内致病力测定,获知该菌致病力差异明显,其中强致病力菌株所占比例大。同时,本研究也对327份苹果种质资源进行了室内抗病性鉴定,其中高抗资源160份,中抗资源6份,中感资源22份,高感资源139份。表明我国现保存的苹果种质资源中存在丰富抗病种质。进一步按苹果分类系统分析发现,抗病资源在当前栽培的主要品种群中均有分布,特别是红玉品种群、富士品种群抗病资源最为丰富。     

覆盖材料和沟垄比对土壤水分和紫花苜蓿干草产量的影响

为寻求半干旱黄土高原区种植紫花苜蓿的适宜覆盖材料和最佳沟垄比,采用完全随机设计布置大田试验,以传统平作为对照,研究不同垄覆盖材料(土壤结皮、生物可降解地膜和普通地膜)和不同沟垄比(沟宽:垄宽分别为60∶30、60∶45和60∶60,单位是cm)对土壤水分和紫花苜蓿干草产量等的影响。结果表明:通过对2012年和2013年紫花苜蓿生育期降雨量统计,2a平均值显示,无效降雨次数(53次)大于有效降雨次数(27次),无效降雨对总降雨量的贡献率(19%)小于有效降雨(81%)。就紫花苜蓿全生育期而言,与平作相比,SR_(30)、SR_(45)、SR_(60)、BMR_(30)、BMR_(45)、BMR_(60)、CMR_(30)、CMR_(45)和CMR_(60)(SR、BMR和CMR分别代表土垄、生物可降解膜垄和普通膜垄,下标分别表示垄宽为30、45cm和60cm)连续2a的平均根层(0—140 cm)土壤贮水量分别提高12.8、19.2、24.4、26.0、30.7、40.5、29.9、37.1 mm和47.7 mm。垄沟集雨种植第1年龄和第2年龄紫花苜蓿根层没有出现明显干层。与平作相比,SR_(30)、SR_(45)和SR_(60)的连续2a紫花苜蓿平均实际干草产量分别降低3%、8%和13%,WUE分别提高52%、58%和55%;BMR_(30)、BMR_(45)、BMR_(60)、CMR_(30)、CMR_(45)和CMR_(60)的连续2a紫花苜蓿平均实际干草产量分别提高14%、12%、7%、17%、19%和9%,WUE分别提高49%、62%、59%、51%、67%和56%。当紫花苜蓿生育期降雨量为380.7—427.6 mm和沟垄比为60 cm∶35—36 cm时,生物可降解膜垄和普通膜垄的紫花苜蓿实际干草产量达到最大值,为该地区垄沟集雨种植紫花苜蓿提供参考。     

人工释放苜蓿切叶蜂的营巢行为观察

苜蓿切叶蜂Megachile rotundata Fabricius是紫花苜蓿的理想授粉昆虫,国内外已广泛利用切叶蜂为紫花苜蓿授粉,因此,对切叶蜂的田间活动行为分析具有重要的实践指导意义。对苜蓿切叶蜂的田间营巢行为进行观测,结果表明:整个花期切叶蜂均十分活跃,放蜂的第4天开始适应环境,正常活动,7月8～22日,切叶蜂最为活跃,进入蜂巢的切叶蜂数最高时达426头/蜂巢,每日10:00～16:00为活动高峰。每日每块蜂巢的封孔数最多为14个,平均每个蜂室封闭巢孔的数量最多为185个。     

紫花苜蓿F-box蛋白基因MsFTL的克隆及功能分析

FTL(F-box Triple LRR protein)是F-box蛋白家族的成员,具有F-box保守结构域,在植物抵御逆境胁迫过程中起重要作用。本研究参考低温胁迫下紫花苜蓿转录组数据设计引物,通过RT-PCR克隆获得紫花苜蓿MsFTL基因,该基因的全长1422 bp,编码473个氨基酸。该蛋白含有1个F-box结构域及3个LRR重复。系统进化分析表明,MsFTL与蒺藜苜蓿XP_003626345.1 F-box/FBD/LRR-repeat protein亲缘关系最近。两者蛋白序列比对发现共有11个差异位点。在低温、盐、干旱以及外源ABA处理下,MsFTL基因受到诱导,表达量上调。构建植物过表达载体pCBM-MsFTL,通过农杆菌介导法转化烟草。对经过抗性筛选、PCR和Real-time PCR验证的转基因植株进行低温抗性鉴定。在-4℃低温胁迫下,野生型烟草叶片出现了明显的萎蔫失水现象,而转基因烟草萎蔫程度相对较轻。生理检测结果表明,4℃处理24 h之后,转基因烟草的可溶性蛋白含量、可溶性糖含量、SOD活性,CAT活性高于野生型,MDA含量低于野生型。本研究表明,MsFTL基因在提高植物对低温胁迫的抗性方面具有重要的作用。     

镉胁迫对紫花苜蓿幼苗生理特性和镉富集的影响

以"甘农三号"紫花苜蓿幼苗为材料,在水培条件下,探究了在10 d内不同浓度(0～2.0 mmol·L^-1)镉(Cd)胁迫对其根长、茎长、生物量、叶绿素和丙二醛(MDA)含量、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性、Cd富集及其亚细胞分布的影响。结果表明:低浓度(0.125 mmol·L^-1) Cd能促进幼苗根和茎的生长,增加叶片叶绿素含量,较高浓度(0.5～2.0 mmol·L^-1) Cd显著抑制幼苗根和茎的生长,叶绿素及生物量显著降低; Cd胁迫使MDA含量显著增加,而SOD和POD活性显著增强,其中Cd胁迫浓度为0.5mmol·L^-1时,SOD和POD活性达到最大值,这可能是植物对环境胁迫的一种应激保护反应; Cd在各亚细胞组分中的含量依次为细胞壁>细胞质>线粒体>叶绿体,且均随Cd胁迫浓度的升高而增加。当Cd胁迫浓度为0.125 mmol·L^-1时,水培10 d的紫花苜蓿幼苗单株地上部对Cd的净化率最高可达0.214%,而整盆植株在单位体积内对Cd的净化率最高可达15.5%;当Cd胁迫浓度为2.0 mmol·L^-1时,紫花苜蓿幼苗地上部Cd含量达89.36μg·g-1。这些结果表明紫花苜蓿对Cd具有很强的富集能力,虽未达到Cd超富集植物的临界标准,但从植株生物量、耐Cd能力、富集Cd量及对Cd的净化率等方面综合考虑,紫花苜蓿在Cd污染土壤的植物修复中具备良好的应用价值。