Identification and characterization of partial resistance to rust in a germplasm collection of Lathyrus sativus L.

Lathyrus sativus has high potential both as food and fodder crop but can be severely infected by rust. This study assessed the resistance level of a collection of L. sativus accessions against three rust species ( Uromyces pisi , U. ciceris-arietini and U. viciae-fabae ) and characterized, for the first time, macro and microscopically the resistance to U. pisi . All L. sativus accessions were highly resistant to U. ciceris-arietini and to U. viciae-fabae but showed a compatible interaction with U. pisi. However, in spite of this compatible interaction, high levels of rate reducing resistance were identified with significant disease severity reduction, fitting the Partial Resistance definition. Detailed histological observations showed that resistance was mainly because of haustorium formation restriction with high-level of colony early abortion, and reduction in the haustoria number per colony. This was occasionally associated with reduction on the infection hyphae intercellular growth, in a clear example of prehaustorial resistance. Differences in germination, orientated germ tube growth and appressoria differentiation were observed but were, generally, of marginal importance to explain the observed resistance levels.     

豆科牧草对滨海盐渍土壤盐分特性和肥力影响的研究

[目的]探讨豆科牧草用于滨海盐渍土生物改良的可性行。[方法]利用田间长期定位试验,研究种植豆科牧草林生山黧豆和紫花苜蓿的人工草地对滨海盐渍土壤盐分特性和肥力的影响。[结果]与自然盐碱空白地相比,长期种植紫花苜蓿和林生山黧豆的人工草地的土壤盐分含量明显降低,0～40 cm土层脱盐最为强烈,脱盐率达47.8%～48.8%,土壤盐分分布由柱状型转变为底积型;0～20 cm土壤pH值明显降低;0～20 cm土层的有机质、全氮含量分别提高了28.4%、42.5%和95.6%、69.8%。紫花苜蓿草地与自然盐碱空白地的盐分离子运移存在着差异。[结论]中度盐渍化土壤种植紫花苜蓿,轻度盐渍化土壤种植林生山黧豆,是生物改良滨海盐渍土的有效途径。     

山黧豆CASase基因DNA全长序列的扩增及CRISPR/Cas9敲除载体的构建

【目的】克隆山黧豆β-腈基丙氨酸合成酶基因(CASase)DNA全长序列,构建CRISPR/Cas9敲除载体,为筛选"低毒、高硫"山黧豆品系奠定基础。【方法】以萌发4d山黧豆幼苗根部为材料,提取其基因组DNA,利用PCR克隆CASase基因全长;经序列同源性、内含子、外显子、sgRNA靶位点和脱靶分析后,在CASasecDNA上选取5个sgRNA靶位点进行寡核苷酸链设计和体外活性检测;从5个sgRNA靶位点中取活性较高的4个sgRNA,利用酶切连接法构建基因敲除载体pPLHACas9-sgRNA20r、pPLHACas9-sgRNA68r、pPLHACas9-sgRNA225f和pPLHACas9-sgRNA256f。【结果】获得了CASase基因3 143bp的DNA全长序列,该序列编码β-腈基丙氨酸合成酶;在CASase cDNA中选取5个sgRNA靶位点在体外均能有效剪切靶序列;菌落PCR检测结果显示,4个CRISPR/Cas9基因敲除载体构建成功。【结论】克隆了山黧豆CASase基因DNA全长,成功构建了该基因上4个sgRNA靶位点的CRISPR/Cas9敲除载体。     

H2O2胁迫对山黧豆生理生化指标的影响

用不同浓度H_2O_2的水培液处理7d山黧豆幼苗24h,分析山黧豆叶片鲜质量、H_2O_2和O_2~-·组织化学定位、丙二醛(MDA)与抗坏血酸(ASA)质量摩尔浓度和过氧化氢酶(CAT)、过氧化物酶(POD)及抗坏血酸过氧化物酶(APX)的活性变化,研究H_2O_2诱导氧化胁迫下山黧豆叶片的生理应答特征,揭示山黧豆叶片对氧化胁迫的耐受机制。结果显示:与对照组相比,5mmol·L~(-1)和10mmol·L~(-1) H_2O_2处理组叶片鲜质量无显著变化,20mmol·L~(-1) H_2O_2处理叶片鲜质量显著降低。山黧豆叶绿素a、叶绿素b和类胡萝卜素的质量分数并无显著变化。随外源H_2O_2处理浓度不断增加,叶片组织内源O_2~-·染色范围和程度呈明显减少趋势,H_2O_2染色范围和程度呈先增加后减少趋势。H_2O_2与MDA积累水平也呈现先增加后减少的趋势;山黧豆叶片中CAT、POD与APX的活性均表现先升高后降低的趋势,而ASA质量摩尔浓度则呈现先降低后上升的趋势。研究表明,叶片内H_2O_2和MDA水平变化与抗氧化酶的活性变化均处于动态平衡中并呈现一致变化趋势,低浓度H_2O_2处理可以提高山黧豆抗氧化酶的活性。     

Assessment of Sensitivity to Frost in Ochrus chickling (Lathyrus ochrus (L.) D.C.) by Chlorophyll Fluorescence Analyses

Ochrus chickling ( Lathyrus ochrus (L.) D.C.) is a promising feed legume for rain-fed farming systems in the Mediterranean environments of West Asia and North Africa (WANA) because of its drought tolerance and resistance to broom rape ( Orobanche crenata Forsk.). However, it is highly susceptible to cold and improving Its resistance would further improve its adaptation.
The potential of the chlorophyll fluorescence technique in screening for frost sensitivity in seedlings of five Lathyrus ochrus genotypes and one each of Vicia ervilia (L.) Wild and V. sativa L. was assessed by measuring the rate of rise in induced or variable chlorophyll fluorescence of fully developed leaflets after cold hardening in the open and subsequent exposure to freezing. The method differentiated the highly frost-susceptible L. ochrus genotypes from the more tolerant Vicia species. Testing the technique with nine genotypes of L. ochrus in which the plants were established, cold-hardened and frost-stressed under controlled conditions confirmed its sensitivity even to reveal the narrow inter plant differences that generally exist within a cultivar or genotype to frost. It is therefore considered as a useful screening technique for cold/frost tolerance in L. ochrus.     

五脉山黧豆开花动态及有性繁育系统的研究

观测了五脉山黧豆的开花动态,并用杂交指数(OCI)、花粉-胚珠比(P/O)方法测定了其有性繁育系统类型。结果表明,五脉山黧豆的花期持续45 d左右,盛花期为6月下旬左右。开花的日动态呈明显的双峰现象,2峰分别在8:00-11:00和14:00-15:00时出现。五脉山黧豆的杂交指数(OCI)=3,繁育系统为自交亲和,有时需要传粉者;花粉-胚珠比(P/O)=2 286.21,为兼性异交。得出其繁育类型为混交繁育系统,即自交和异交并存。五脉山黧豆的传粉系统不完善和虫害是结实率低的主要原因。     

Flatpea crossing methods and germplasm screening for reduced 2,4-diaminobutyric acid composition

Summary Flatpea (Lathyrus sylvestris L.) is a potentially valuable forage legume, but it contains high levels of 2,4-diaminobutyric acid (A₂bu), a compound that can have adverse effects on some animals, including rats and poultry. This study evaluated intercrossed pollination methodology needed for cultivar improvement and characterized intercrossed progeny for A₂bu concentration. Pollen germination on an artificial medium, using pollen collected during different stages of flower development, was used to identify appropriate stages for flower emasculation. Pollen dehiscence occurred at an early-bud stage when petal color was first evident. Slightly more mature buds contained pollen with substantially higher in vitro germination than pollen from early-bud stage flowers. Lines that produced high numbers of seeds per pollination were identified from crosses between flatpea accessions. Cross-pollination resulted in significantly higher seed set than selfing. Pollination using caged honeybees was the most efficient method of hybridization, although the degree of inbreeding in bee-pollinated plants could not be determined. Intercrossed progeny produced by honeybee pollination contained a lower mean concentration of foliar A₂bu than the mean for flatpea accessions.Abbreviations HPLC high performance liquid chromatography - OPA ortho-phthalaldehyde     

New approaches towards the shortening of generation cycles for faster breeding of protein legumeshh

In order to shorten generation cycles, a greenhouse strategy was used as control and compared with an in vitro plus in vivo strategy for pea and bambara groundnut, as well as to an in vitro only strategy for pea and grass pea. Using an in vitro plus in vivo system and embryo axis explants, nearly six generations per year for Pisum and over four generations for Vigna were obtained, compared with two generations in the field. Using successive generations from seed to seed in pea, the mean duration for one generation was 67.2 ± 4.6 days in ‘Frisson’, against a mean of 143 ± 3 days in the field. With the in vitro only strategy in pea, 6.87 generations per year were obtained with ‘Frisson’ and 5.24 with ‘Terese’, while with grass pea genotypes over three generations per year were possible. All plants obtained were morphologically normal and fertile. These results show the feasibility of using such strategies to reduce significantly the duration of generation cycles in legumes, thus offering novel approaches for breeding these important crops.     

Diversity of seed albumins in some Lathyrus species related to L. sativus L.: An electrophoretic study

Electrophoretic analysis of seed albumins covered 66 Lathyrus accessions representing L. amphicarpos, L. blepharicarpus, L. cicera, L. gorgoni, L. marmoratus, L. pseudocicera and L. stenophyllus. The number of major albumin bands recorded in particular species varied from two in L. amphicarpos to eight in L. gorgoni. Apart from L. amphicarpos, all the species under study showed intraspecific variation. In total, 21 bands were distinguished. The results were compared with the corresponding data reported earlier for L. sativus. All the data were statistically analysed. The species examined in this study proved to be distantly related to L. sativus. Distinctness of particular Lathyrus species was quite marked except for L. cicera and L. marmoratus which produced similar electrophoretic seed albumin patterns.