A water-soluble polysaccharide, designated as KOMAP, was isolated and purified from the alkaline extract of king oyster mushroom, which was composed of glucose (Glc), mannose (Man) and arabinose (Ara) in a molar ratio of 6.2:2.1:2.0. It had an average molecular weight of 2.1×10(4)Da. GC-MS analysis revealed that KOMAP was a linear structure of the polymer with a backbone composed of β-1,4-linked glucopyranosyl and β-1,3,6-linked mannopyranosyl units, which was terminated with α-1-linked arabinofuranosyl unit at C-6 position of β-1,3,6-linked mannopyranosyl residue along the main chain in the ratio of 3.1:1. The results in the animal experiment showed that 50, 100 and 200mg/mL of KOMAP not only inhibited the tumor growth, but also increased relative thymus and spleen indices, LPS- or ConA-induced lymphocytes proliferation, and serum cytokine IL-2, TNF-α, and IFN-γ levels, as well as the activities of NK cells and CTLs in spleen of Renca tumor-bearing mice. In summary, our data indicate that the KOMAP exerts effective immunoregulatory and anti-tumor activities in vivo.Copyright © 2014 Elsevier Ltd. All rights reserved.

对原生质体融合技术研究进展进行了综述。从原生质体融合的步骤——原生质体制备、培养及融合入手，分别详细阐述了原生质体制备、培养和融合等方面的研究进展，分析了影响原生质体融合的相关因素，随后对原生质体融合的方法、方式以及融合获得的杂种细胞的筛选、鉴定进行了概述，并结合有关文献资料，综述了杂种细胞常用的筛选及鉴定方法，最后对这一技术在育种实践中存在的问题进行了分析，并对其今后的应用前景进行了展望，为原生质体融合技术的发展提供了参考。

Volvariella volvacea is difficult to store fresh because of the lack of low-temperature resistance. Many traditional mutagenic strategies have been applied in order to select out strains resistant to low temperature, but few commercially efficient strains have been produced. In order to break through the bottleneck of traditional breeding and significantly improve low-temperature resistance of the edible fungus V. volvacea, strains resistant to low temperature were constructed by genome shuffling. The optimum conditions of V. volvacea strain mutation, protoplast regeneration, and fusion were determined. After protoplasts were treated with 1% (v/v) ethylmethylsulfonate (EMS), 40 Sec of ultraviolet (UV) irradiation, 600 Gy electron beam implantation, and 750 Gy Co-γ irradiation, separately, the lethality was within 70%-80%, which favored generating protoplasts being used in following forward mutation. Under these conditions, 16 strains of V. volvacea mutated by EMS, electron beam, UV irradiation, and Co-γ irradiation were obtained. The 16 mutated protoplasts were selected to serve as the shuffling pool based on their excellent low-temperature resistance. After four rounds of genome shuffling and low-temperature resistance testing, three strains (VF, VF, and VF ) with high genetic stability were screened. VF, VF, and VF significantly enhanced fruit body shelf life to 20, 28, and 28 H at 10 °C, respectively, which exceeded 25%, 75%, and 75%, respectively, compared with the storage time of V23, the most low-temperature-resistant strain. Genome shuffling greatly improved the low-temperature resistance of V. volvacea, and shortened the course of screening required to generate desirable strains. To our knowledge, this is the first paper to apply genome shuffling to breeding new varieties of mushroom, and offers a new approach for breeding edible fungi with optimized phenotype.© 2015 International Union of Biochemistry and Molecular Biology, Inc.

【目的】了解不同食（药）用菌原生质体分离与再生情况,为原生质体诱变及融合提供材料,从而有目的地选育有突出优良性状的新菌株。【方法】以5种食（药）用菌菌丝体为试验材料,利用酶解法制备5种食（药）用菌原生质体。【结果】在相同条件下,原生质体释放量：桑黄zsh>草原白蘑BM1716>杏鲍菇xb>草原黑蘑hm8124-1>蟹味菇xwg;再生率：蟹味菇xwg>杏鲍菇xb>草原黑蘑hm8124-1>草原白蘑BM1716>桑黄zsh。进一步观察草原黑蘑hm8124-1的原生质体再生单个菌落生长情况发现,其菌丝更加洁白、粗壮、浓密;跟踪观察6个草原白蘑BM1716原生质体再生单个菌落生长情况则出现明显差异,菌落的生长形态和生长速度不同,出现角变现象。【结论】不同食（药）用菌品种之间原生质体的释放量与再生率存在很大差异,需在后续试验中优化再生培养基。6个草原白蘑BM1716原生质体再生单个菌落生长差异非常明显,并且出现了角变现象,很有可能在原生质体分离与再生过程中发生了体细胞变异。