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.     

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.     

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。     

不同集雨种植方式对干旱区紫花苜蓿种植的影响

在甘肃省定西半干旱黄土丘陵区采用完全随机设计,研究不同覆盖材料(普通地膜CMR、生物可降解地膜BMR和土垄SR)和不同沟垄比(60 cm∶30 cm、60 cm∶45 cm和60 cm∶60 cm)对径流效率、土壤贮水量、土壤含水量、紫花苜蓿干草产量和水分利用效率的影响.结果表明：SR、BMR和CMR的平均径流效率分别为32.0%、90.7%和96.4%.在紫花苜蓿生育前期(4-6月),各处理之间土壤贮水量差异不显著;在紫花苜蓿生育后期(7-10月),BMR和CMR的土壤贮水量显著高于SR,SR的土壤贮水量显著高于平作,如在紫花苜蓿孕蕾期,平作、SR、BMR和CMR 0~140 cm土层土壤贮水量分别为223.27、248.56、277.81和284.16 mm.平作、SR、BMR和CMR全生育期实际干草产量分别为4112.1、3397.5、4317.8和4523.8 kg·hm^-2,水分利用效率分别为11.08、10.48、14.56、14.95 kg·mm^-1·hm^-2.在同一覆盖材料下,不同沟垄比对土壤贮水量和水分利用效率的影响不显著.当沟垄比为60 cm∶44 cm时,CMR和BMR处理的实际产量均达到最大.     

Predation drives stable coexistence ratios between red and green pea aphid morphs

We conducted field surveys and experiments to evaluate the hypothesis that predation is an important driving factor determining the degree of coexistence between red and green morphs of the pea aphid Acyrthosiphon pisum. Theory suggests that the different colour morphs are differentially susceptible to natural enemies and selection by predation which in turn leads to variable relative abundances of red and green morphs among host plants across landscapes. Our field surveys on pea and alfalfa revealed, however, that the colour morphs tended to coexist closely in a ratio of one red to three green aphids across fields with different host plant monocultures. Experimentation involving manipulation of the relative abundances of the two colour morphs on host plants pea and alfalfa with and without predator presence revealed that red morphs had higher or same fitness (per capita reproduction) than green morphs on both pea and alfalfa only when in the proportion of one red/three green proportion. Moreover, experimentation evaluating predator efficiency revealed that red morphs are safest from predation when in a 1 : 3 ratio with green morphs. These results suggest that in addition to predation selection effects, red morphs may behaviourally choose to associate with green morphs in a narrow 1 : 3 ratio to maximize their fitness. This evidence, along with existing published data on red and green morph anti‐predator behaviour indicates that a 1 : 3 red and green morph coexistence ratio is driven by a balance between predation pressure and behavioural assorting by red morphs across landscapes. In this way predators may have ecological‐evolutionary consequences for traits that affect the colour morphs' proportion and tolerances to selective pressure.     

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。     

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。