IJM - Italian Journal of Mycology ISSN 2531-7342 - Vol. 50 (2021): 63-77 Journal homepage: https://italianmycology.unibo.it/ Review The cultivation of Lactarius with edible mushrooms Di Wang1, Jun Li Zhang2, Yun Wang3, Alessandra Zambonelli4, Ian R. Hall5*, Wei-Ping Xiong5 1 Institute of Agricultural Resources and Environment, Sichuan Academy of Agricultural Sciences, 4# Shizishan Road, Chengdu, Sichuan, People’s Republic of China 610066 2 Tibet Academy of Agricultural and Animal Sciences 147 West Jingzhu Road, Lhasa, Tibet, People’s Republic of China 850032 3 Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan, People’s Republic of China 650204 4 Department of Agricultural Science, University of Bologna, Via Fanin 44, 40127 Bologna, Italy 5 Truffles and Mushrooms (Consulting) Ltd, P.O. Box 268, Dunedin, New Zealand 9054 * Corresponding author e-mail: [email protected] - The arrangement of the authors is simply alphabetical first for the name of their counties followed by an alphabetical arrangement for their given names. ARTICLE INFO Received 13/5/2021; accepted 10/6/2021 https://doi.org/10.6092/issn.2531-7342/12908 Abstract From the early 1800s science virtually ignored the cultivation of edible mycorrhizal mushrooms other than the true truffles. The drought was finally broken by Nicole Poitou when she cultivatedLactarius deliciosus and Suillus granulatus in the Institut National de la Recherche Agronomique’s laboratories at Pont-de-la-Maye, France, in the mid-1970s. However, another 20 years were to pass before Yun Wang working at Invermay Agricultural Centre near Dunedin, New Zealand, was able to begin the routine cultivation of L. deliciosus. The purpose of this paper is to document the history of the cultivation of L. deliciosus in New Zealand since its introduction in the late 1990s, its spread back to Europe and its subsequent introduction into Yunnan, Tibet and Sichuan starting about 2014. Keywords saffron milk cap, cultivation, New Zealand, China, productivity Introduction The past 50 years has seen several revolutions in the way that plants have been produced for the cultivation of edible mycorrhizal mushrooms. The first method for truffles commonly referred to as “Talon’s Technique” (Hall and Zambonelli, 2012; Hall et al., 2007), dates back to the early 1800s and involves growing clean seedlings in the rooting zone of plants already mycorrhized with truffle or in soil known to have produced truffle in the past (Rocchia, 1995). However, the collapse in truffle production particularly in France between 1900 and 1960 (Hall and Zambonelli, 2012) led to calls for alternative methods for producing truffle mycorrhized plants. Three methods were forthcoming. That described by Palenzona (1969) for Tuber melanosporum Vittad., Tuber aestivum Vittad., and Tuber brumale Vittad. involved inoculating each plant with about 107 ascospores from blended truffles. Another method replaced spores with sections of root already mycorrhized with truffle (Chevalier and Grente, 1973) a method independently developed for inoculating plants with arbuscular mycorrhizal fungi (Hall, 1973, 1976). A third method employed pure cultures to inoculate clean seedlings Department of Agricultural and Food Science in partnership with the Italian Mycological Society (UMI). The journal is hosted and mantained by ABIS-AlmaDL. © 2021 The Authors. This work is licensed under CC BY 4.0 Wang et al. Italian Journal of Mycology 50 (2021): 63-77 https://doi.org/10.6092/issn.2531-7342/12908 (Chevalier 1972, 1973; Chevalier and Grente, 1973; Fontana and Palenzona, 1969). But by the end of the 1970s it was the first of these techniques that was to be generally adopted for the cultivation of truffles in Europe and by Hall in New Zealand in the mid-1980s (Hall et al., 2007; Murat, 2015). Sporal inocula have also been used in plantation forestry when it was found essential when plants were raised in soilless media in greenhouses, sufficient inocula could not be expected to blow in from surrounding vegetation, and when outplant sites were devoid of suitable ectomycorrhizal fungi. For example, in New Zealand a suspension of blended Rhizopogon roseolus (Corda) Th. Fr. in water is often sprayed onto plants in containerised Pinus radiata nurseries (Hall et al., 2003a; Visnovsky et al., 2010) and Rhizopogon parksii A.H. Sm. onto Douglas fir [Pseudotsuga menziesii (Mirb.) Franco] (Hall et al., 2019). Because ascospores and basidiospores are produced sexually there is the expectation that spores from, for example, Tuber borchii Vittad. truffles from the northern reaches of this species’ distribution, say near Edinburgh, are likely to be better adapted to the climatic conditions there than T. borchii from Sicily. So, when Christina Wedén successfully grew T. melanosporum on Gotland off the east coast of Sweden, way north of its natural distribution, it was with plants inoculated with truffles harvested from the northernmost distribution of T. melanosporum in France (Wedén et al., 2013). By the mid-1990s demand for truffle mycorrhized plants in New Zealand had increased and considerable income was being made from their production and sale by Crop & Food Research (Hall et al., 2007). Despite the commercial revenue from this, research funding was more or less reduced in line. So, Hall’s group needed new scientific and hopefully commercial challenges and decided to import Lactarius deliciosus (L.) Gray (saffron milk cap) (Figs 1a, b, c and d). This has an excellent flavour, international market appeal, and importantly, there was some pioneering work that had been published in France by Nicole Poitou and Jacques Delmas that goes back to the mid-1970s showing that it was possible to culture the fungus and get it to fruit in the field (Delmas, 1978; Poitou, 1978, 1982; Poitou et al., 1981, 1984, 1989). Our overlying logic for carrying out this work was because L. deliciosus is a mycorrhizal fungus there was the possibility of growing it as a secondary crop in New Zealand’s 1.6 million hectares of Pinus radiata D. Don forest (Berg, 2008). Also, because it forms mycorrhizas exclusively with pines, there was little chance that it would enter New Zealand’s native ectomycorrhizal beech forests and compete with the local mycorrhizal fungal flora, we were given permission by New Zealand Biosecurity to import it. In retrospect, perhaps we should have considered trying to import L. sanguifluus and L. vinosus as well which are more popular than L. deliciosus in some countries. In this paper we outline the initial introduction of L. deliciosus into New Zealand, subsequent production of its fruiting bodies, the expansion of research aimed at commercial production, and the establishment of several lines of research that emanated from those early days in Tibet, Sichuan and Yunnan. Whilst L. deliciosus is the species that ranks at the top of a list of the most desirable species alongside porcini (Boletus edulis Bull.: Fr sensu lato) and chanterelles (Cantharellus spp.), elsewhere in the world there are other species of Lactarius rated equally highly (Supplementary Table 1). Consequently, a commercial success with L. deliciosus on P. radiata was likely to create interest with other species of Lactarius and on other hosts elsewhere in the world as later outlined by Leonardi et al. (2017). We explore the literature on the edible species of Lactarius in the hope that it will shed light on the methods that have been used for their cultivation, what impediments have hobbled their production on commercial scales and, in particular, their cultivation out of season from the Northern Hemisphere like truffles are now being grown in the Southern Hemisphere (Hall et al. 2017a). Hopefully it might also shed light, in particular, on the cultivation of edible Lactarius throughout China. The techniques we describe here have worked for us but no doubt methods described by others may have some advantages (e.g. Poitou et al. 1984; Díaz et al. 2009; Wang et al. 2012, 2020). 64 Wang et al. Italian Journal of Mycology 50 (2021): 63-77 https://doi.org/10.6092/issn.2531-7342/12908 Fig 1 - Fruiting bodies of Lactaris deliciosus (a, b, c and d). Some of the first saffron milk mushrooms to be harvested by Hannes and Theres Krumenacher near Nelson, New Zealand (a) and by John and Portia Barnes from a Pinus radiata plantation near Castlemaine, Victoria, Australia (b) Bleeding an orange-coloured sap when the caps or stalks are damaged (c) which become green when exposed to the air (d). Hunan street vendor selling Lactarius hatsudake, a popular edible species particularly in China, Japan, Korea and eastern Russia (e). The poisonous brown roll rim (Paxillus involutus) (f) has been mistaken for the saffron milk cap but it has a different colour, it does not bleed an orange-coloured sap when damaged, it turns reddish-brown when cut and has a tightly involuted margin to the cap (g). In New Zealand, a country where mycophagy (mushroom eating) is not strong, the poisonous brown roll rim (Paxillus involutus (Batsch) Fr., Figs. 1f and g) has been mistaken for the saffron milk cap. There are also some poisonous Lactarius lookalikes such as the woolly milk cap (Lactarius torminosus (Schaeff.) Gray) (Hall et al., 2003b) and there are photos on the internet which show there is some justification for making such a mistake (e.g. https://en.wikipedia.org/wiki/Lactarius_torminosus). In China, plants raised in greenhouses and inoculated with Lactarius sect. Deliciosi (a section of the genus Lactarius) can be heavily infested by dipterous larvae which render their mushrooms 65 Wang et al. Italian Journal of Mycology 50 (2021): 63-77 https://doi.org/10.6092/issn.2531-7342/12908 inedible (Ran et al., 2019). It has been suggested that this is because the antifeedant properties of the mild-tasting latex of the group are relatively ineffective (Ran et al., 2019; Nuytinck, 2004-2005).