层出镰刀菌乌头酸酶家族的功能研究

【目的】层出镰刀菌是引起苜蓿根腐病的主要病原菌之一,探究层出镰刀菌中乌头酸酶家族蛋白的功能特性,为深入认识层出镰刀菌基础生理代谢的分子机制提供依据。【方法】利用hmmsearch工具,对真菌中含有乌头酸酶结构域的蛋白进行检索,并进行系统进化分析;通过实时荧光定量PCR及SWISS MODEL建模技术分别分析FpACO基因的表达模式与蛋白结构;利用同源重组双交换方法构建层出镰刀菌乌头酸酶基因敲除突变体;分析ΔFpACO3、ΔFpACO4-1、ΔFpACO4-2敲除突变体的生长、产孢、孢子形态、环境胁迫响应及致病力等表型变化;进一步测定敲除突变体中线粒体代谢相关生理生化指标的变化情况。【结果】FpACO4-1与FpACO4-2在产孢及孢子形态发生中发挥作用;FpACO3、FpACO4-1、FpACO4-2参与调控层出镰刀菌对细胞壁胁迫及金属离子胁迫的敏感性;FpACO3、FpACO4-1、FpACO4-2影响线粒体代谢,包括总乌头酸酶活性、ATP含量、过氧化氢含量及三羧酸循环关键基因表达等。【结论】乌头酸酶家族参与调控层出镰刀菌产孢、孢子形态分化、细胞壁胁迫及金属离子胁迫响应和线粒体代谢等过程。     

Detection and Identification of Two Phytoplasmas (16SrIII‐B and 16SrXII‐A) From Alfalfa (Medicago sativa) in Serbia

Plants of alfalfa (Medicago sativa) exhibiting general stunting, proliferation and phyllody associated with leaf yellowing and reddening were observed in three localities of Central Serbia. Phytoplasma strains belonging to 16SrIII‐B and 16SrXII‐A groups were detected and identified by RFLP and sequence analysis of 16S rDNA. Stolbur phytoplasma tuf gene RFLP analysis showed the presence of the TufAY‐b‐type phytoplasma subgroup in 80% of symptomatic samples. This is the first report of 16SrIII‐B and 16SrXII‐A phytoplasma groups affecting alfalfa in Serbia.     

Embryoid formation in alfalfa cell suspension cultures from different plants

Summary Cell suspension cultures were initiated from shoot tips of nine plants of alfalfa. Our results indicated that considerably different nutritional conditions were required to induce embryogenesis in the cell cultures derived from these plants. By proper adjustment of the hormone level and mineral salt concentration it was possible to induce embryogenesis in all of the nine cultures tested. The embryoids from seven of the nine cultures were able to develop into plants.     

Resistant Host Responses to Ten California Populations of Meloidogyne incognita

Resistant and susceptible cultivars of tomato, lima beans, cotton, and alfalfa were tested with 10 populations of Meloidogyne incognita from different California locations. Nine of the populations differed in aggressiveness on the nine cultivars tested. Two populations were especially aggressive toward resistant tomato cultivars.     

A simple and effective method for isolating RNA from alfalfa pollen

Extraction of RNA from alfalfa pollen using conventional grinding devices resulted in low yields of degraded RNA; degradation appeared to be related to the length of time required to break open the pollen grains with such devices. A glass syringe was modified to provide the restricted, shearing environment necessary for rapidly rupturing this type of tissue. This apparatus would be useful for extracting a variety of molecules from pollen of any species, but is particularly valuable for species which produce low amounts of relatively small pollen grains.     

Performance of the tobacco aphid Myzus persicae (Hemiptera: Aphididae) on Oriental and Virginia tobacco varieties

Abstract 1 The life parameters of one red and one green clone of Myzus persicae reared on 11 Oriental tobacco type varieties (KP7, K63, KZ10/z, MA13/b, BX2/a, N34/4, KP14/a, Kolindros, S79, TA21 and BX81) and on five Virginia ones (VE9, VE3, SPG28‐VE2, Kleio‐KN1 and Niki‐NK3) at 20 °C and LD 16 : 8 h were examined. 2 In general, the clones, performed better on Oriental than on Virginia type varieties. The varieties TA21 and Kolindros could be considered as the most resistant among the Oriental ones. Aphids on Virginia varieties showed longer developmental time (9.1–9.6 days), a lower intrinsic rate of increase (0.2347–0.2647) and higher nymphal mortality (27.9–52.5%) than on Oriental varieties. The corresponding values in the latter were 7.3–9.0 days, 0.2784–0.3302 and 10.0–47.9%. 3 The leaves of Virginia type varieties (188–471 hairs/cm²) and one Oriental (K63) 273 hairs/cm² were more hairy than the other Oriental type varieties (61–150 hairs/cm²). The Virginia type varieties showed also a lower level of reducing sugars (3.5–7.2%) than the Oriental ones (7.0–9.8%). Trichome density and level of reducing sugars were significantly negatively correlated with developmental time and intrinsic rate of increase. 4 An interclonal variation in performance, possibly related to the colour of aphids, was observed. The red clone performed better than the green one. The same trend was observed when the results of the previous clones on the variety KP14/a were analysed with the data obtained on the same variety from two other clones (one red and one green).