Nodulation of lucerne (Medicago sativa L.) in an acid soil: Effects of inoculum size and lime-pelleting

The effects of inoculum level and lime-pelleting were studied in an acid soil with respect to the nodulation and growth of lucerne (Medicago sativa cv Resis) and the population dynamics of Rhizobium meliloti. In small root-boxes (rhizotrons), the in-situ survival of inoculated rhizobia was studied in the micro-environment around the seed for a period of 12 days after sowing. During the initial 24 hours, a strong increase in rhizobial numbers was measured, concomitantly with the development of roots. As a result of lime-pelleting, rhizobial numbers were higher only at 3 days after sowing (P<0.05). Later, this difference diminished steadily. Addition of lime did not increase the adhesion of the rhizobia to the seedling tap root. Plant responses to inoculation were studied in pots. To obtain optimal nodulation, the soil had to be neutralized around the seed with lime and at least 10⁵ cells of R. meliloti were required. With more than 10⁵ rhizobia per seed, lime-pelleting increased the number of crown-nodulated seedlings from 24% to 77%. Higher numbers of rhizobia could not compensate the effect of lime. A strong correlation was found between crown nodulation, nitrogen content and dry weight of the shoots.     

Survival of inoculated Laccaria bicolor in competition with native ectomycorrhizal fungi and effects on the growth of outplanted Douglasfir seedlings

The survival, development and mycorrhizal efficiency of a selected strain of Laccaria bicolor along with naturally occurring ectomycorrhizal fungi in a young plantation of Douglas fir was examined. Symbionts were identified and their respective colonization abilities were determined. Eight species of symbiotic fungi, which may have originated in adjacent coniferous forests, were observed on the root systems. Mycorrhizal diversity differed between inoculated (5 taxa) and control (8 taxa) seedlings. Ectomycorrhizal fungi which occurred naturally in the nursery on control seedlings (Thelephora terrestris and Suillus sp.) did not survive after outplanting. Both inoculated and naturally occurring Laccaria species, as well as Cenococcum geophilum, survived on the old roots and colonized the newly formed roots, limiting the colonization by other naturally occurring fungi. Other fungi, such as Paxillus involutus, Scleroderma citrinum and Hebeloma sp. preferentially colonized the old roots near the seedling's collar. Russulaceae were found mainly in the middle section of the root system. Mycorrhizal colonization by Laccaria species on inoculated seedlings (54%) was significantly greater than on controls (13%) which were consequently dominated by the native fungi. Significant differences (up to 239%) were found in the growth of inoculated seedlings, especially in root and shoot weight, which developed mainly during the second year after outplanting. Seedling growth varied with the species of mycorrhizae and with the degree of root colonization. Competitiveness and effectiveness of the introduced strain on improving growth performances of seedlings are discussed.     

HBsAg合成肽段的免疫源性和抗肽血清与天然HBsAg的结合测定

我们将三种乙型肝炎表面抗原的合成肽段在联结载体或不接载体的情况下,分别制备成4种免疫原对兔子进行了免疫。大部分兔子都产生了抗肽抗体。其中抗P_(122-48)(auw)与抗P_(122-148)(adw)都能与天然HBsAg反应,而抗P_(122-148)(adw)的抗HBs活力比文献报道的都高,通过结构分析表明:含有天然蛋白上免疫显性区域的免疫原是合成疫苗的理想选者。     

百日咳杆菌去内毒素及保护性抗原的纯化

内毒素(Endotoxin,简称ET)是百日咳全菌苗(Bordetellapertussis vaccine)产生副作用的主要毒素之一,且不易除去。现有的分离方法,如蔗糖密度梯度离心法,较繁琐,成本高。本文采用Sepha-cryl S-300凝胶层折法可以简便有效的去除大部分内毒素。初步毒素试验结果表明:已达到日本生物制品规格的要求。两种保护性抗原FHA和LPF-HA也得到进一步分离纯化,为今后研制高效的百日咳组分菌苗提供了实验条件。     

烟草内生菌根真菌的分离鉴定

本文报道了从烟草(Nicotiana tabacum L.)的根际土壤中分离出内生菌根真菌的孢子,用单孢接种烟苗并在温室内水培条件下培养。选出能在烟草上形成菌根的菌株,经过再次单孢接种确认后,进行种的鉴定。从分离的8个菌株中已鉴定出球囊霉属(Glomus)的3个新记录种:漏斗孢球囊霉[G.mosseae(Nic.& Gerd.)GeM.& Trappe]、根内孢球囊霉(G.intraradics Schenck & Smith)和联结球囊霉(G.constrictum Trappe)。     

Experimental Histomoniasis in the Guinea Fowl,Numida meleagris*

Guinea fowl were inoculated rectally with Histomonas meleagridis to produce histomoniasis. The birds were infected readily by this unnatural route. Severe cecal involvement was frequent and long-lasting but liver lesions and death were rare. Turkeys given the same inoculum had high levels of liver involvement and mortality. Guinea fowl responded much less severely to infection with H. meleagridis when infected by a natural route (ingestion of a vector) rather than by rectal inoculation. Thus, naturally acquired infections with H. meleagridis appear to be of small consequence in the guinea fowl.     

Nitrogen fixation and growth of soybean as influenced by varying the methods of inoculation with Bradyrhizobium japonicum

The effects of inoculating soil with a water suspension of Bradyrhizobium japonicum (i) at seeding, (ii) 7, or (iii) 14 days after planting (DAP), (iv) seed slurry inoculation and (v) seed slurry supplemented with postemergence inoculation of a water suspension of Bradyrhizobium at 7 or (vi) 14 DAP, on nodulation, N₂ fixation and yield of soybean (Glycine max. [L.] Merrill) were compared in the greenhouse. The ¹⁵N isotope dilution technique was used to quantify N₂ fixed at flowering, early pod filling and physiological maturity stages (36, 52 and 70 DAP, respectively). On average, the water suspension inoculation formed the greatest number of nodules, and seed plus postemergence inoculation formed slightly more nodules than the seed-only inoculated plants (27, 19 and 12 nodules/plant respectively at physiological maturity). Seed slurry inoculation followed by postemergence inoculation at 14 DAP gave the highest nodule weight, with the plants fixing significantly more (P<0.05) N₂ (125 mg N plant⁻¹ or 56% N) than any other treatment (mean, 75 mg plant⁻¹ or 35% N). However, the higher N₂ fixation was not translated into higher N or dry matter yields. Estimates of N₂ fixed by the ostemergence Bradyrhizobium inoculations as well as plant yield were not significantly different from those of the seed slurry inoculation. Thus, delaying inoculation (e.g., by two weeks as in this study) did not reduce the symbiotic ability of soybean plants.     

表达HSV－2 gD基因的重组痘苗病毒保护小鼠对抗致死量HSV病霉攻击

利用ＰＣＲ方法对单纯疱疹病毒Ⅱ型糖蛋白Ｄ（ＨＳＶ－２ｇＤ）基因进行了修饰,在其５＇端删去约５００ｂｐ的非编码区,仅保留ＡＴＧ上游７个ｂｐ。将修饰后的ＨＳＶ－２ｇＤ基因插入到带有痘苗病毒天坛株ＴＫ基因区段的痘苗表达质粒ｐＪＳＡ１１７５,置于痘苗病毒Ｐ７．５ｋ早／晚期启动子控制下。将此重组质粒用脂质体Ｌｉｐｏｆｅｃｔｉｎ方法转染已受野型ＴＫ￣＋痘苗病毒天坛株感染的ＴＫ￣－１４３细胞,通过同源重组机制和标志基因ＬａｃＺ产物的蓝斑显色作用,以及ＢｕｄＲ试剂对ＴＫ表型的选择压力,筛选出整合有ＨＳＶ－２ｇＤ基因的重组痘苗病毒。Ｓｏｕｔｈｅｍ杂交表明,ＨＳＶ－２ｇＤ基因已正确地插入痘苗病毒ＴＫ基因区内;间接免疫荧光检测显示,ＨＳＶ－２ｇＤ蛋白已得到有效表达,且主要分布于细胞膜。重组病毒免疫家兔可产生明显的抗ＨＳＶ－２ｇＤ中和抗体。用重组病毒免疫小鼠,３周后可使９４％（１７／１８）的小鼠对抗ＨＳＶ－２的致死量攻击,表明重组病毒具有明显的免疫保护作用。     

Growth and yield of lentil and wheat inoculated with three Glomus isolates from Saskatchewan soils

The vesicular-arbuscular mycorrhizal fungi (VAMF) Glomus clarum (Nicol. and Schenck) isolate NT4, G. mosseae (Nicol. and Gerd.) Gerd. and Trappe isolate NT6 and G. versiforme (Karst.) Berch isolate NT7 coexist in wheat field soils in Saskatchewan. This study assessed the response of lentil (Lens esculenta L.) and wheat (Triticum aestivum L.) to monospecific and mixed cultures of these VAMF isolates. Seedlings were inoculated with 100 spores of a VAMF isolate, or an equal mixture of spores of two isolates, and grown in a sterile soil mix in a growth chamber. Both crops responded differently to these different VAMF isolates. In the case of lentil, G. clarum NT4 was more effective than G. mosseae NT6 and G. versiforme NT7, and significantly increased (P<0.05) the shoot dry weight (43%) and grain yield (57%) compared with the uninoculated control. There was a significant positive correlation between the percentage of VAMF colonized roots and shoot dry weight (r=0.672^***) and shoot phosphorus concentration (r=0.608^***) of lentil. In the case of wheat, G. clarum NT4 had no effect on shoot dry weight, but produced significant (P<0.08) increases in grain yield (12%) and the phosphorus concentration of the shoot and grain. Although G. clarum NT4 and G. mosseae NT6 both produced similar levels of VAM colonization in wheat, the only response of wheat to isolate NT6 was an increase in plant height at harvest. The efficacy of G. clarum NT4 on both crops appeared to be related to its ability to produce more arbuscular colonization than G. mosseae NT6. Dual inoculation of seedlings with G. clarum NT4 and G. mosseae NT6 resulted in competition between these two isolates. This was evident from a comparison of plant shoot dry weight and grain yield, and VAMF spore production on the two crops inoculated either with isolate NT4 alone or in combination with NT6. G. mosseae NT6 reduced the efficacy of G. clarum NT4 by 16% when dual inoculated on lentil, but had no effect when the host was wheat. Based on spore production, it was found that G. clarum NT4 was more competitive than G. mosseae NT6 when dual inoculated on lentil or wheat. Isolate NT4 produced ca. 2000 and 500 spores/ 100 g substrate, respectively, in the lentil and wheat pots, which was approximately 2–3 times more spores than those produced by isolate NT6 with either crop. When the plants were dual inoculated, there was a 15–19% reduction in spore production by G. clarum NT4 and a 50–70% decrease in spore production by G. mosseae NT6. Our results show that G. clarum NT4 was more competitive and effective in its ability to colonize and increase the growth and yield of lentil and wheat than G. mosseae NT6 or G. versiforme NT7. The relative performance of isolate NT4 with different host plants suggests that this VAMF isolate exhibits a host preference for lentil.