Report of Italian VIR/IHA Cannabis Germplasm Preservation Project - 1995

Alexandra Doubovskaja1, Andrea Sonnino3 , Ugo De Corato2 , Lyudmila Rumyantseva1, and Robert C. Clarke4

1 N. I. Vavilov Institute of Plant Industry, 42 Bolshaya Morskaya Street, 190000 St. Petersburg, Russia; Institute for New Technology, Energy and the Environment: 2 Trisaia Station, S. S. Jonica 106, 75025 Policoro (Matera), Italy and 3 Casaccia Station, 301 Anguillarese Street, 00060 S. Maria di Galeria (Rome), Italy; 4 International Hemp Association, Postbus 75007, 1070AA Amsterdam, The Netherlands

    Doubovskaja, A., A. Sonnino, U. De Corato, L. Rumyantseva and R. Clarke, 1996.  Report of Italian VIR/IHA Cannabis Germplasm Preservation Project - 1995.  Journal of the International Hemp Association 3(1): 7-9.
    Fourteen accessions of Cannabis from the germplasm collection of the Vavilov Institute were sown at 7 locations in Italy in 1995 for germplasm maintenance and reproduction of seed reserves for distribution.  Ten accessions yielded from 73 to 7,500 grams of seeds.   The other 4 yielded no seeds.  The plant pathogens Phomopsis sp. and Macrophomina phaseolina (Tassi) Goid. were reported on the hemp plants near Policoro.

    In 1993, the N. I. Vavilov Institute of Plant Industry (VIR), with the support of the International Hemp Association (IHA), began a program to preserve its Cannabis germplasm collection.  In 1994 the project was expanded to the Ukraine and Italy. Accessions of Italian origin were selected for sowing in Italy in order to reproduce them in the most suitable conditions.  Another goal of this project was to return to Italy its own hemp germplasm to conserve it not only in the VIR Gene Bank, but in Italian Gene Banks also.   The VIR collection contains 8 Italian Cannabis accessions (Lemeshev et al. 1994), and most of them were of low viability.  All Italian accessions in threatened condition were sown in Italy in 1995.
    Accessions of a Chinese Cannabis group with long vegetative cycles were also chosen for sowing in Italy in 1995.  It is difficult to reproduce them in northern Russia and the dissolution of the Soviet Union caused the separation of some southern VIR stations outside Russia.

Table 1.  Participants of the VIR/IHA Cannabis Germplasm Preservation Project in Italy - 1995.
  Responsible Organizations VIR Catalog No.
  Experimental Institute of Industrial Crops of Bologna, Dr. Paolo Ranalli
University of Bologna, University of Milan, Prof. Maria T. Amaducci
Peasant Civilization Museum of Bologna, Mr. Ivano Trigari
General Agriculture and Grass Crops Inst., Sassari, Prof. Giuseppe Rivoira
Institute for New Technology, Energy and the Environment:
               Casaccia Station of Rome, Dr. Andrea Sonnino
               Trisaia Station of Policoro, Dr. Alexandra Doubovskaja
278, 462
280, 282
184, 336

181, 279, 323, 340
112, 182, 321

Materials and methods
    Fourteen accessions of hemp from the VIR Germplasm collection were sown at 6 locations in Italy (Table 1).   This material was introduced to the collection from Italy, China, Turkey and Bulgaria, and represents varieties, lines and local populations (Table 2).  For the purpose of germplasm maintenance three collection accessions with the origin from Italy and China were sown in two plots at the Trisaia Station of the Institute for New Technology, Energy and the Environment (ENEA) near Policoro.  The distance between the plots was 850 meters and contained trees and high buildings.  At one plot were planted a landrace accession from Italy (n.112) and 'Chain-Chgo' from China (n.182) with a difference in vegetative cycles of 22 days.  The Italian early-ripening accession was sown 10 days before the Chinese late-ripening variety to avoid stray pollination.   Shared plantings of temporally isolated varieties allows twice as many reproductions in the same amount of space.  The Chinese accession Shan-Ma (n. 321) was planted at the second plot.
    The first accession was sown on May 22.  The seeds were sown in densely planted rows spaced 30 cm apart.  The size of the plot varied from 5-8 m2 depending on the number of seeds. The plots were irrigated after sowing and later on depending on plant conditions.  The soil was predominantly composed of red clay.
    The other 7 accessions were sown in order to look for the possible correlation between the contents of tetrahydrocannabinol (THC) and some clearly visible early developmental morphological markers.  Five accessions with high-THC from the IHA, and variety 'Yellow stem' and line 'Foglia Pinnatofida' from VIR collection were investigated; but they did not show the yellow color of the stem and the pinnate type of leaf respectively.  Six from the seven accessions were taken away at the phase of 6-8 pairs of leaves, before the first male flowers opened.  The variety 'Yellow stem' was left at the plot to look for the stem color at a later phase of vegetation.
    Four accessions were sown at the Casaccia Station of ENEA near Rome on two double plots: the Italian 'Bolognese' (n.279, vegetative cycle 140 days) together with the Chinese 'Sen-Ma' (n.323 166 days), and the Bulgarian 'J-3' (n.340 140 days) with the Chinese 'Sar-San' (n.181 190 days).  The sowing method was similar of that at the Trisaia Station.
    At the Peasant Civilization Museum of Bologna, the local Italian variety (n.195) was sown on April, 15 on a plot of 16 m2.  The distance to other hemp sowings was 5 km.  The plants were covered with nets at the time of ripening to protect the seeds from birds.  The plots were irrigated twice.  The soil was clayish.
    The accession 'Distretto di Fatza' (n.280) originally from Turkey was sown at the University of Bologna, and the Italian accession 'Carmagnola' (n.282) was sown near Milan.  At the Industrial Crops Institute of Bologna two Italian accessions 'Napoletana' (n. 278) and 'Linea a foglia pinnatofida' (n. 462) were sown on April, 3 and May, 22 respectively.  The distance between the plots was more than 70 km and the size of the plots were 48 and 60 m2.  The seeds were sown in the rows spaced 40 m 2 and 100 cm apart with the distance between the plants in the row 13 and 10 cm.  The soil was clayish.
    Two Chinese accessions 'Tin-Yan' (n.184) and 'Dun-Ma' (n.336) were sown at the General Agriculture and Grass Crops Institute of Sassari on May, 29 and on June, 9 respectively.  The distance between the plots was more than 100 km.  The seeds were sown in densely planted rows spaced 50 cm apart.
    Agrometeorological conditions in Italy during 1995 were favorable for the growth of hemp.  The spring, first half of summer and autumn were hot and dry.   August was humid with frequent rainfall totaling 96 mm compared to the average of 12 mm.  Because of this both early-ripening varieties and late-ripening varieties found good conditions for maturation.
    The number of plants was calculated at the beginning of growth or at complete flowering (total plant number), and before the harvest (female plant number).   The average height of plants for the plot was measured before the harvest.   The vegetative cycle was calculated from the day of sowing to the day of complete maturation and/or harvesting.

  Table 2.   Results of Italian VIR/IHA Cannabis Germplasm Reproduction Project-1995.
[Note: Veg. cycle is missing data that is summarized in the text.]
Name/Origin Total
Local, Italy
Local, Italy
Napoletana, Italy
Bolognese, Italy
Carmagnola, Italy
Linea a foglia pinnatofida, Italy
Distretto di Fatza, Turkey
J-3, Bulgaria
Sar-San, China
Chain-Chgo, China
Tin-Yan, China
Shan-Ma, China
Sen-Ma, China
Dun-Ma, China








    Ten reproductions yielded from 73 to 7,500 grams of seeds.  The largest yields were obtained in the Industrial Crops Institute and in the Peasant Civilization Museum of Bologna from Italian hemp accessions (Table 2).
    Four accessions (ns. 181, 280, 282, 340) yielded no seeds because of the poor germination or the absence of germination (Table 2).  Accession n. 280 did not germinate in spring and was sown again with the remaining seeds.  The plants were transferred into the greenhouse to obtain some seeds during the winter period.
    The vegetative cycle varied from 119 to 176 days for the Italian accessions of hemp and from 115 to 144 days for the Chinese ones.  The plant height varied from 90 to 350 cm for the Italian hemp accessions and from 95 to 195 cm for the Chinese ones.
    The plants of the variety 'Yellow stem' developed different shades of the stem color from green to almost yellow.  The seeds from two plants with the most yellow stem color were harvested for later experiments.
    Two fungal diseases were identified on the plants at the ENEA Trisaia Station near Policoro.  The "bad withering" disease became apparent as complete withering of the plant at the beginning of maturation.  The seeds of infected plants were visibly normal and they ripened normally; 15-20 days earlier than the seeds of the healthy plants.  Fourteen percent of the plants were infected in the plot of accession n. 321.  There were black microsclerotia on the roots of infected plants (Figure 1).  According to Ferri (1959) it was Macrophomina phaseolina (Tassi) Goid., which is usually transmitted by vegetative remains.
    Another disease became apparent as numerous little black picnidia on the stem around the branch.  The zone of infection was clearly distinct from the healthy tissue (Figure 2A) or it had no clear border (Figure 2B).  The conidia of the fungi were unicellular with two nuclei, suggesting a classification as Phomopsis sp.. (Von Arx, 1981).   This infection is usually transmitted by water (Goidanich, 1975).  There are no data about the possibility of transmission of these two diseases by the seeds.


Figure 1.  Microsclerotia of Macrophomina phaseolina (Tassi) Goid. on the root of the hemp plant at the ENEA Trisaia station in 1995 (Photo G. Mazzei). Figure 2.  The picnidia of Phomopsis sp. on the stem of the hemp plant at the ENEA Trisaia station in 1995.  A - the zone of infection is clearly located, B - the zone of infection is without border (Photo G. Mazzei).

    In the first report of the VIR/IHA Cannabis Germplasm Preservation Program two key requirement were defined to consider an accession adequately reproduced: at least 1000 plants have to produce at least 200 g of seed (Lemeshev et al. 1994).
    Due to poor germination only 7 from 10 sown accessions reproduced sufficient quantity of seed.  Among these 7 accessions, only one had a sufficient population size.
    All reproduced seeds of the Chinese accessions and a part of the seed of each Italian accession have been sent to the gene bank of the VIR.  The remainder of the seed of each Italian reproduction will be preserved in Italy.
    A sample of seeds of the Italian industrial hemp variety "Carmagnola" has been received by the National Flax and Hemp Industry of Vimercate (Milan).  Part of the seeds will be preserved in Italy.  The remaining seeds have been sent to the VIR.

    This project was possible, thanks to support from the International Hemp Association and the participation of Prof. Maria Teresa Amaducci, Prof. Alessandro Bozzini, Dr. Giancarlo Messagi, Dr. Paolo Ranalli, Prof. Giuseppe Rivoira and Mr. Ivano Trigari.