How 2016/17 black truffle season has gone in France & Spain?


import - export black truffle in France, marketing and prices for black truffle

In France producers and traders estimate for the 2016/17 harvest the volume of French truffle production to represent 30 tonnes of Tuber melanosporum and 25 tonnes of Tuber aestivum var uncinatum and mesentericum. A half-tinted campaign (44.8 tons for 2015/16), which took place more favorably in irrigated truffles.
The federation of nursery/truffle tree producers and the regional federations of truffle growers estimated the professional plantations at 1,200 hectares for the year 2016.

FranceAgrimer just published a great document with lots of interesting data on truffle market, prices and volumes, exports and imports:

http://www.franceagrimer.fr/content/download/51148/491610/file/Truffe%20-Présentation%20Commerce%20extérieur%20au%2010052017.pdf

The truffle production during the last season was bad in Spain, in fact, there were no truffle at all on the wild in most regions, however, is the first season that I can remember – from the past 20 years that I´ve been working with truffles- that even being such a bad season, total quantities (wild + farms) harvested in Spain were higher than last year. This is because every year we have more and more plantations with irrigation. Prices were higher in average than other years so growers (who could water) were quite happy.

Prices paid at harvester (dirty and all qualities mixed) in the largest black truffle market (Teruel) has been between 400-750€/kg.  In Catalonia every year more restaurants use truffles in their menus, this made that local truffle harvesters start selling their best truffles directly to local restaurants and leave the worst truffles to be sold at the weekend truffle market. This has lead to drop prices at local markets (due to low quality) and even some traders are not interested to drive to these markets to buy anymore, so some markets are disappearing. This is of course a pity as we loose a tradition but at the same time means that truffle consumption is growing in Spain and this is something to congratulate.

Cheers,

Marcos S. Morcillo

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Where and when males come from to fertilize truffles?


two-year-old-oak-producing-its-first-black-truffle

In the following paper we hypothesize about a new theory we created related with black truffle life cycle and when spores germinate, if they do it as males or females. We are working in order to prove it in coming research… I would love to hear your comments…

Cheers,

Marcos Morcillo and Xavier Vilanova

________________________________________________________________________

New considerations of Tuber melanosporum life cycle

Xavier Vilanova1, Marcos Morcillo1, Mónica Sánchez1, Herminia De la Varga1

1 Micologia Forestal & Aplicada. Camí d’Alfou sn, 08459 Sant Antoni de Vilamajor, Barcelona, Spain.

Corresponding author: xaviervilanova@micofora.com

 

Introduction

The black truffle, Tuber melanosporum Vittad., is an ascomycete fungi that produces ectomycorrhizal associations with different trees and shrubs (Mello et al. 2006). The black Périgord truffle is known by the organoleptic properties of their fructifications, that are naturally harvested under mediterranean regions, but also cultivated all over the world in orchards planted with inoculated seedlings. Since 2015 it is considered as a perennial culture for the European Union, although the production of inoculated seedlings is done since the 70s (Murat 2015), and thus the importance of study the species. After the publication of T. melanosporum genome (Martin et al. 2010), in the last years a huge progress has been done to unravel truffle life cycle (Le Tacon et al. 2016, Taschen et al. 2016, De la Varga et al. 2017). Nowadays we have more information about how the fungus live, in symbiosis with plants, and in soil. We have information coming from scientific research, from experimental truffle sites, wild forests and established truffle plantations.

Regarding research publications, we have an approximation to T. melanosporum life cycle. It is a heterotallic specie, mating is controlled by two idiomorphs called MAT 1 and MAT2 (Rubini et al. 2011). Truffles are the result of sexual reproduction, we know that a female individual (that can be either MAT 1 or MAT 2), found always as mycorrhiza, has to meet with a male one of the opposite mating type (whose origin can be different). Most of individuals acting as fathers come from germinating ascospores, as its principal function is the sexual reproduction (De la Varga et al. 2017, Taschen et al. 2016).

In different publications, it has been described that it exists, at root level, a kind of progressive exclusion of the female individuals harboring the mating type opposite to the dominant one, leading to have plantations with a patchy distribution of the mating types at root level, having each tree roots with individuals of only one mating type (Murat et al. 2013, De la Varga et al. 2017).

One of the practices that is widespread among truffle growers, specially in Spain, is to make the called “truffle traps”. This technique consists on doing holes or trenches around the trees and to introduce a substrate with a mixture of spores. In those substrates, the spores come from a mixture of different truffles, so a mixture of both mating types. In a recent publication (Murat et al. 2016) it was tested the efficiency of this technique and showed how truffle production could be promoted and localized in a few centimeters, opening also the possibility to better understand the role of ascospores.

Discussion

In our experiences along the years, when we observe the behavior of the spores throughout the different steps in the process of truffle production in managed plantations (plant production, plantation establishment and management techniques – substrate addition to increase truffle production – ) we have seen that:

  • For the production of melanosporum mycorrhized seedlings, the inoculation takes place in march-april (in the northern hemisphere) observing the first mycorrhizae in October-November (observing the roots under a microscope). Moreover, using molecular techniques, as the Real-Time PCR (Parladé et al. 2013) it can be detected an increase of germinated mycelia, coming from the spores, from July. (unpublished data)
  • Periodically, inoculated seedlings are analyzed to check the level of mycorrhization of the roots. When doing those analyses in nursery plants, but also in two years old trees, that have been planted in the field, non-germinated spores are detected. They remain in the substrate, near the roots, and apparently, they are viable spores.
  • As it has been noticed in field experiences (data not shown) and in other publications (Murat et al. 2016), in the places where substrate has been applied truffles fructify 2 years after the application. The results of adding spores to the soil surrounding the trees are reflected as an increase of the production of truffle ascocarps, that are localized in the place where the substrate has been applied (Murat et al. 2016).

All those evidences make us hypothesize that the first germinating spores form the ectomycorrhiza, so that they could be “predetermined” to act as mother/female individuals, or they “choose” to germinate and act as mother/female elements and form the ectomycorrhizae. The other spores remain in the soil waiting to germinate next year or later. This could also explains why the results of applying substrates with spores are not seen after 2 years. Indeed, it has been reported the fructification of truffles in plantations with inoculated trees of two years old (data not shown). In this case, it could be explained by the germination of the spores that remained in the substrate, and that they act as male elements for the sexual reproduction.

This simple hypothesis could explain the behavior observed in the spores at the different stages of truffle life cycle. Currently, the mechanisms that determine that one individual acts as female or male element are unknown (genetics, environmental, molecular, etc.), but probably this function is determined. It could be that in the genome of each individual it was determined so that the first spores to germinate act as female element, that could entail an energy saving for the fungus, as well as an explanation to the rates of consanguinity described in some publications (Taschen et al. 2016).

The fact that T. melanosporum fructifies in traps with added spores after 2 years, and in young two years old trees planted in orchards, could be explained by this hypothesis. Both kind of fructifications could be promoted by the late germination of the spores that will act as male elements. In the example of truffle traps these male partners will come from the spores added to the substrate; for the young seedlings, they may come from the bank of spores remaining in the substrate of the plant. The question is that if this is like that, we should expect to have fructification in older plants maintained in nurseries. This has never been reported, probably the germination of the ascospores that will act as male elements is triggered by environmental signals and that is why they remain non-germinated as a bank of spores.

Conclusion

The last years research has advanced to unravel how is the life cycle of the black truffle (Murat et al. 2013, Le Tacon et al. 2016, Taschen et al. 2016 and De la Varga et al. 2017), but there are still some points to be clarified. More research needs to be done to explain how and why one individual act as female or male element, especially how this is regulated. Those findings would have an impact in truffle industry, as methodologies to promote mating will lets us to have more control on the production of truffle ascocarps.

References

De la Varga, H., Le Tacon, F., Lagoguet, M., Todesco, F., Varga, T., Miquel, I., Barry-Etienne, D., Robin, C., Halkett, F., Martin, F. and Murat, C. (2017), Five years investigation of female and male genotypes in Périgord black truffle (Tuber melanosporum Vittad.) revealed contrasted reproduction strategies. Environmental Microbiology. Accepted Author Manuscript. doi:10.1111/1462-2920.13735

Le Tacon, F., Rubini, A., Murat, C., Riccioni, C., Robin, C., Belfiori, B., Zeller, B., De la Varga, H., Akroume, E., Deveau, A., Martin, F., Paolocci, F. (2016) Certainties and uncertainties about the life cycle of the Périgord black truffle (Tuber melanosporum Vittad.). Annals of Forest Science, 73: 105. doi:10.1007/s13595-015-0461-1

Martin, F., Kohler, A., Murat, C., Balestrini, R., Coutinho, P.M., Jaillon, O., et al. (2010) Perigord black truffle genome uncovers evolutionary origins and mechanisms of symbiosis. Nature. 464(7291):1033–8.

Mello, A., Murat, C., Bonfante, P. (2006). Truffles: much more than a prized and local fungal delicacy. FEMS Microbiology Letters 260, 1–8.

Murat, C. (2015) Forty years of inoculating seedlings with truffle fungi: past and future perspectives. Mycorrhiza 25, 77–81.

Murat, C., Rubini, A., Riccioni, C., De la Varga, H., Akroume, E., Belfiori, B., Guaragno, M., Le Tacon, F., Robin, C., Halkett, F., Martin, F. and Paolocci, F. (2013), Fine-scale spatial genetic structure of the black truffle (Tuber melanosporum) investigated with neutral microsatellites and functional mating type genes. New Phytologist, 199: 176–187. doi:10.1111/nph.12264

Murat, C., Bonneau, L., De la Varga, H., Olivier, J.M., Sandrine, F. and Le Tacon, F. (2016) Trapping truffle production in holes: a promising technique for improving production and unravelling truffle life cycle. Italian Journal of Mycology 45. doi: 10.6082/issn.2531-7342/6346

Rubini, A., Belfiori, B., Riccioni, C., Tisserant, E., Arcioni, S., Martin, F., and Paolocci, F. (2011) Isolation and characterization of MAT genes in the symbiotic ascomycete Tuber melanosporum. New Phytologist 189: 710 722.

Taschen, E., Rousset, F., Sauve, M., Benoit, L., Dubois, M.-P., Richard, F. and Selosse, M.-A. (2016), How the truffle got its mate: insights from genetic structure in spontaneous and planted Mediterranean populations of Tuber melanosporum. Molecular Ecology, 25: 5611–5627. doi:10.1111/mec.1386

 

Here a link for the pdf of this paper: https://www.slideshare.net/micofora/new-considerations-of-tuber-melanosporum-life-cycle-1

 

 

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how to decide a layout for truffle farming in the USA?


The last two weeks we have been visiting 8 truffle farming projects in the USA with @xvilanovasola, mostly in California and in North Carolina. Let me comment some of the issues we mostly find and to compare the climates and soils.

In Ashville (NC) is probably where we find temperatures more similar to the ones in Catalonia. i.e. 23,7ºC as average of the hottest month (July) and 2,8ºC as average in the coldest month (january). Here black truffle will enjoy direct sunlight to the ground, so better make layouts with rows North-South with low density (150 trees/acre).

Ashville 2ashville 1

In this area it rains all year round, with 70-80mm every month, so some orchards would not even need irrigation during fruiting stage.

On the other hand, in California temperatures are quite higher, specially the average of the maximum in the hottest month. The amplitude of temperatures during day & night are huge, so truffle will probably look for some shade in some areas, so we can design layouts a little more dense or with rows East-West.

sonoma climate truffles micoforaSonoma valley truffle growing Morcillo

Another issue that may raise is when truffle season ends in mid march, we have to do all tasks in a truffle orchard, this is to prune, chop branches, add truffle spores or spanish wells and rip or till before spring starts. But in CA most trees already move/sprout in late march, we also found mycorrhizae were active in most samples in early april. This means that the “window” to do more tasks is shorter than in Spain. 2 weeks in CA while 4-5 weeks in Spain. And in north California rains in march so soil may be too wet to work before spring comes…

Here two pics one with old Quercus suber and another with Q. robur & pubescens with black truffle. The first with an excessive canopy and the second will close soon if trees are not heavily pruned:

quercus suber produces trufflesquercus robur mycorrhized with black truffle micofora

Rain is one of the main problems in the area north of San Francisco, like Napa and Sonoma Valley, specially in winter where the amount of water is pretty high at the time that truffle are harvested. This will create problems in the orchards in the valleys, with clay soils and bad drainage. Ripping or improving drainage (french drains) will be a must to avoid truffle rot.

We found several orchards with excessive tree density that need to be thinned, trees toped or heavily pruned:

hazel mycorrhized with black truffle micoforaXavier Vilanova samples a filbert truffle orchard

Cheers,

Marcos S. Morcillo & Xavier Vilanova

 

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Truffle Growing in México


truffle growing in México

I attach a picture of the 1300 Pinus pinea that we planted in Zumpango, México in 2015 and their evolution since planted.
In another pilot plot more to the north we have 1 hectare with 250 stone pines plus another with Pinus cembroides. This other pine also produces edible nuts. In the same plantation we have some Mexican native oaks mycorrhized as well.

The mycorrhizal levels of the samples sent to our lab in Barcelona are very good and we will try this spring to quantify the concentration of mycelium of Borchii with the molecular technique Real Time PCR.
pino micorrizado borchii México
We are considering the possibility to graft these pines with highly productive clones of pine cones, to shorten the waiting time to pine cones production and increase nuts yield.
Another key factor for these plantations to enter into production will be to add special substrates with bianchetto spores to provide the “male factor” that can fertilize the maternal part found in the soil and mycorrhizae, as we do with the spanish wells with the black truffle.
Piñonero Zumpango trufa Méjico
These pilot plantations will be the first to produce truffles and European pine nuts jointly, two non-timber products of high market value.

If you want to know more about the reproduction of this truffle:

Characterization of the reproductive mode and life cycle of the whitish truffle T. borchii

Cheers from California, where my partner Xavier Vilanova and I are visiting several truffle projects. Will keep you posted!

Note the next International Workshop on Edible Mycorrhizal Mushrooms  IWEMM9 will be this summer in Mexico

 Marcos S. Morcillo

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Truffle Farming worldwide update for 2017


how many hectares of truffles are planted in the world

A spanish newspaper (el Diario de Teruel) asked me some data to create an updated paper and Infographic about truffle farming in the world, and this is how it looks like, although it is in spanish I though it was nice to show you.

Rechecking now I can detect some edition mistakes, i.e first truffle plantation in Australia was on ´92 and they got the first truffle in Tasmania in ´99…

They made some interviews as well to the growers that attended the last edition of the Barcelona Truffle Tour, from USA, Australia, South Africa or Israel among others.

Cheers,

Marcos S. Morcillo

P.D. I´ve been extremely busy this truffle season, but I will keep on writing in this blog soon 🙂 as there are lots of ideas and papers to show here…
 

 

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Some bacteria and active organic compounds boost truffles mycelium


We tested 8 bacteria strains + 6 organic compounds on nursery and truffle plantations

My partner Dr. Xavier Vilanova just gave a lecture about the new results from a joint research project we do with IRTA in Barcelona.

As you´ll see in the embed lecture we tested 8 bacteria strains and 6 organic compounds and quantify 3 months later the concentration of black truffle mycelium on each trial.

Main conclusions are:

• Each stage (age) of the plant has a different reaction for each treatment.

• In nursery phase, 11 of the 14 treatments generated a positive effect on the

concentration of mycelium. Mainly two organic substances (TR11 and

TR12) and a bacterial strain (TR9).

• In plantation of 3.5 years old: all treatments generated a positive effect on the

concentration of truffle mycelium in the soil. Mainly two organic substances

(TR14 and TR15) and two bacterial strains (I TR9 TR6). These trees have started as well to produce truffles while the others don´t. So there is a significative correlation between the treatment, the concentration of truffle mycelium and later black truffle fruitings.

• In 10 years old plantation 5 to 14 treatments generated a positive effect on the

concentration of truffle mycelium in the soil, all organic substances

(TR10,11,13,14 I15).

Bellow you can see a picture of one tree in the trial farm, where last week we just found a black truffle under a just 2 year old Quercus ilex

two-year-old-oak-producing-its-first-black-truffle

Cheers,

Marcos S. Morcillo & Xavier Vilanova

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New results from our research with “spanish wells” with black truffle spores


how-to-make-spanish-wells-for-black-truffle-micofora

Last week we have organized with the IRTA (Institute of Research in Agronomic Technologies) a workshop to show the results of our latests join research on truffles.

I embed here my lecture with the results. We quantified black truffle mycelium after “spanish wells” inoculation, with different spore dosis and at different months.

Main conclusions are:

. Drip irrigation system seems not to favour truffle mycelium expansion in soil. 3 logarithms below outside humidity bubble.

• Seasonal differences in distribution of black truffle mycelium in the soil. Seasonal pattern dependent from water or irrigation.

• Significative more Tuber mycelium concentration and sequences in productive trees.

• Idem for Scleroderma. Could it be a positive partner specie?

• April inoculations increase Tuber sp. sequences and mycelium concentration in june, as compared to controls non-inoculated, reaching same levels of sequences than when inoculated in june at 5grams. BUT for agronomic reasons it should be better to inoculate in april, as we mostly do mechanically with tractors over the brulê, in order not to disturb the active mycelium in june.

This is an example of a tool that can be adapted for truffle farming:

. We detect significative more mycelium 2 months after reinoculations, probably due to sporal germination. The main problem nowadays is that we normally make the “spanish wells” with truffle spores right after the truffle season in ended (in norther hemisphere from 15th march) and sometimes the soil is too wet because it rains in spring, so some growers delay this task to avoid compacting the ground. This makes that when truffle spores germinate is too late for a “male mycelium” to do fecundation, and this is why most “spanish wells” produce truffles 2 winters later. Although this is just a theory that need to be proven. We are working on that 😉

A way could be to add truffle spores to the soil before the end of march, but in productive orchards we cannot do it with a tractor as we would damage truffles already in the soil. This is why some growers start adding substrates with spores in each hole where they found a truffle…

Cheers,

Marcos S. Morcillo

 

 

 

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New standards to market truffles in Europe


truffle-grading-qualities

It has just been published the new standards to market and quality control of fresh truffles, not including truffles to be processed for industry.

You can download the full document in the next link:

NORME CEE-ONU FFV-53 concernant la commercialisation et le contrôle de la qualité commerciale des TRUFFES

 

truffle-trees-and-cold

let me give you some updates about truffle season in Spain, which is the worst in decades, with almost no wild truffle in the woods. As always we don´t really know why, we come from the driest winter, so trees hardly sprout last spring and moreover spring and early summer was so warm…who knows…

If you want to keep updated with prices and volumens, follow my partner Dr. Xavier Vilanova every monday´s twit https://twitter.com/xvilanovasola/status/810746170037432320

Quantities are low and in large areas in Catalonia fog has stayed for over a month with continued temperatures around 0ºC. This has lead to damage shallow truffles and not sure how trees will be affected as fog is still there.

Cheers,

Marcos S. Morcillo

 

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Pines will produce Lactarius fruit bodies inside our nurseries!


saffron milk cap mushroom cultivated

Lactarius deliciosus fruiting in a pot from a mycorrhized pine seedling

We are happy these Christmas as we´ll be starting a new research project. This is the mycorrhization of pines with an specific Lactarius delicious strain who produces fruit bodies inside our nurseries, right on the pots!

This strain has been isolated by the team at IRTA and the project have been granted with a Torres Quevedo fund from the spanish Government for 3 years. Here a couple of pics of the mycelium growing on pure culture:

safrron mil cap mushroom mycelia

We are so happy as well because of this project    a new member is joining our team full time: she is Herminia de la Varga, who comes from the Nancy lab at INRA, in France, and she will coordinate the Lactarius project plus our DNA lab.

If you want to check her latests papers and researches, check her profile at Researchgate.

You can download her PhD Thesis where she developed the firsts quantitative tools with DNA to monitor truffles, porcini and saffron milk cap mushrooms on soil:

Welcome Herminia and Merry Christmas and Happy New Year to everyone!

Marcos S. Morcillo

 

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more light on the reproduction cycle of the black truffle (Tuber melanosporum)


elisa-taschen-truffle-life-cicle

I was preparing a lecture for our growers seminars (Barcelona Truffle Tour) and rechecking this recent study presented at the last conference in Cahors and published in the journal Molecular Ecology and produced by a team of the National Museum of Natural History, with the CNRS and the University of Montpellier, gives more light on the reproduction cycle of the black truffle (Tuber melanosporum).

The french truffle season, like in Spain is pretty bad, mostly to the lack of rain in summer, but probably the dry spring that made trees to sprout weakly. Hunters say it will be one of the worst in years. Moreover in this paper they states that the spontaneous sites represent only 20% of the production.
Truffles results from a fertilization between two very different individuals: the first, large, considered as the mother, is associated with the roots of neighboring trees. It persists from year to year and nourishes the truffle. The second, considered the father, is smaller, annual, and not associated with tree roots. It could result from the germination of a non-dispersed spore, or a kind of male that survives only to fertilize a better installed individual.
The planting of inoculated trees did not change the genetic diversity of black truffle populations compared to wild sites. Thus, genes circulate freely between these environments. In one detail: in plantation, the number of fathers is higher and they are more genetically varied, perhaps because of spore intakes, voluntary or not, during truffle treatments.
There is a geographic variability of genetic diversity in the Languedoc-Roussillon region, which is still preserved in plantations. This suggests that the provenances of the inoculated trees should not be mixed too much if it is desired to preserve local genetic characteristics of the black truffle.

Cheers

Marcos S. Morcillo

References:
Molecular Ecology – How the truffle got its mate: insights from genetic structure in spontaneous and planted Mediterranean populations of Tuber melanosporum, par Taschen E., Rousset F., Sauve.M, Benoit L. Dubois M.-P., Richard F., Selosse M.-A.
DOI: 10.1111/mec.13864
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