DELIVERABLE REPORT - fp7-gratitude.eu · MES 11698 Jamur Amis ... MES 03444 G21 Unknown Commercial strain 18 Pleurotus eryngii MES 12060 Le Lion PE Unknown Commercial strain 19 MES

DELIVERABLE REPORT

Grant Agreement number: 289843

Project acronym: GRATITUDE

Project title: Gains from Losses of Root and Tuber Crops

Funding Scheme: Seventh Framework Programme

Date of latest version of Annex I against

which the assessment will be made:

2011-11-15

Deliverable Number: D 4.1

Deliverable Title: Vegetative growth and mushroom production

on cassava waste

Deliverable leader, organisation, tel and

email:

Anton Sonnenberg, Plant Research

International, Wageningen UR. +31 317

481336. [email protected]

Contributing authors to this deliverable: Johan Baars, Marcel Visser

Name, title and organisation of the

scientific representative of the project's

coordinator:

Dirk Pottier, Scientific Officer, European

Commission - DG for Research and Innovation,

E3: COV2 07/157, B-1049 Brussels, Belgium,

Tel: +32 229 67209;

[email protected]

Project website address: www.fp7-gratitude.eu

http://www.fp7-gratitude.eu/

ii

Table of Contents

Table of Contents ............................................................................................................................. ii

1. Summary ......................................................................................................................................... I

Introduction .................................................................................................................................................. I

2. Key Findings ................................................................................................................................. II

3. Deliverable Objectives .............................................................................................................. II

4. Background ................................................................................................................................... II

5. Methodology ................................................................................................................................. II

Preparation of substrates for testing vegetative growth ...................................................................... II

6. Results ............................................................................................................................................ V

6.1 Tests on vegetative growth of fungal species/strains on cassava waste. ....................... V

6.2 Effects of fungal growth on digestibility by rumen microflora ......................................... IX

6.3 Mushroom production of selected strains on cassava waste ............................................ XI

7. Conclusions ............................................................................................................................... XIII

8. References ................................................................................................................................. XIV

Annes 1 Pictures of mushroom production ....................................................................... XIV

Annex 2 Statistical analysis of yield data ............................................................................ XIV

I

1. Summary

Varieties of three different species of edible fungi have been grown vegetatively on cassava

peels, cassava sticks and a mixture of the two (1 : 1 (w/w). Growth has been estimated by

measuring linear growth rate in tubes containing the different substrates. Very accurate

growth rate could be estimated in this way. The order of growth rate on different substrates

was stalks > a 1:1 mixture of stalks & peels > peels. The order of growth rate for different

species was Pleurotus ostreatus > Pleurotus pulmonarius > Pleurotus eryngii > Lentinula

edodes. Variation between strains within one species was limited. After full colonization,

each strain-substrate combination was further incubated for an extra 2, 4 and 6 weeks.

These samples have been analysed for digestibility by ruminants in a model system showing

that especially cassava sticks (stalks) can be made digestible almost comparable to peels

after pretreatment with fungi of edible mushrooms.

Mushroom production of cassava waste (peels, sticks and 1:1 mixtures of peels and sticks)

were used to produce mushrooms of grey oyster and lung oyster mushrooms. Yield up to

15% biological efficiency (fresh weight mushrooms/fresh weight substrate) were achieved

indicating that waste of cassava only is in principle suitable for the production of edible

mushrooms. Addition of nitrogen rich waste (such as rice or wheat bran) will certainly

increase yields to a level comparable to substrates now used (mainly saw dust)..

Introduction

For a viable business on mushroom production on cassava waste materials (peels and

sticks), fungal strains producing edible fungi must first grow well vegetatively on waste

materials. Subsequently, species/strains that show a good colonization of waste can be

tested for the production of edible mushrooms. Species should be selected that are

commercially available now and thus known to consumers and also easy to cultivate. The

first part of the report deals with the selection of the best mushroom species/strains-

substrate combination for small scale testing of mushroom formation on cassava waste. For

this, 30 mushroom strains representing 4 species have been tested for vegetative growth on

cassava peels, cassava sticks and a mixture of the two (1:1). Four strains from FIIRO Nigeria

were included. A selection of strains were incubation for an additional period (2 and 4

weeks) to see if this increases digestibility and makes cassava waste a better substrate for

animal feed.

A selection of 4 strains, representing 2 species, showing a good colonization of different

waste materials were subsequently selected for mushroom cultivation on a small scale to

evaluate the potentials for mushroom production. Strains were selected that were shown to

produce mushrooms on an elevated temperature (>22 oC), since cooling capacity is limited

especially in African countries and would make cultivation on household level and small

enterprises impossible.

II

2. Key Findings

Mycelia of different varieties of the common oyster mushroom, brown oyster mushroom,

king oyster mushroom and shiitake were grown cassava peels, cassava stalks or a 1:1

mixture of both waste products. In general all species and varieties did grow on all waste

types but growth rate differs per species and waste type, i.e. for waste type: stalks> mixture

stalks/peels and for species: Pleurotus ostreatus > Pleurotus pulmonarius > Pleurotus eryngii

> Lentinula edodes.

Pretreatment of especially cassava sticks with fungi increases the digestibility for animals

considerably as tested in a rumen model. Substrates based on cassava waste only can be

used in principle to produce oyster mushrooms with yield up to 150 grams per kilogram

substrate (biological efficiency of 15%). This is high considering the very low nitrogen

content of the cassava waste based substrates. It is expected that addition of nitrogen rich

waste (like rice and wheat bran) can increase yield top levels comparable to saw dust based

substrates used at this moment.

3. Deliverable Objectives

To develop technologies for making food products from waste which is either in the form of

food products, animal feeds or mushrooms made from composting waste.

4. Background

Fungi are specialist in mineralization of a broad range of waste materials, especially the

most abundant waste on earth, i.e. lignocellulose. A large variety of waste steams can thus

be used to produce new food as edible mushrooms. To make cassava production a more

profitable crop it is, therefore, obvious to test also if waste products generated by cassava

production can be used to generate new food products such as edible mushrooms. When

good starting materials is available (strains and inoculum, the latter also known as spawn),

mushrooms is a low tech business and can be done on a small and large scale. Mushroom

production has thus potentials to for developing countries to generate food on waste and

additional income for small enterprises.

5. Methodology

Preparation of substrates for testing vegetative growth

III

During the first half year of 2013 preliminary work was done on optimization of substrate

composition. For this we used peels (milled and sieved, ca. 2 mm sieve, i.e. MESH 60 or 80),

stalks (milled and sieved, ca. 2 mm sieve, MESH 60 or 80) and a mixture of milled peels and

milled stalks (on a dry weight basis 50/50, w/w). For proper wetting, substrates were

submerged overnight in water. After this, excess water was removed from the substrates by

pressing as much water as possible from the substrate. The wetted substrate was used to fill

glass tubes to a height of 12 cm. Substrate was filled into the tubes in small portions. After

each portion, the substrate was “packed” by inserting a metal rod into the tube until it

rested on the substrate. The metal rod fitted neatly into the tubes and by its weight

produced an evenly packed substrate throughout the full height of the tube (i.e. 12 cm).

After this the tubes were capped and sterilised (1 hr. 121 oC). The three different substrates

were tested for their suitability for colonization by 30 strains listed in Table 1. Strains were

first grown on malt extract agar. After a few days of growth (colony diameter about 4-5 cm),

5 mm (Ø) plugs were taken from the edge of the growing colony. In total 5 of these plugs

MES-number Strain name Origin Remarks

Lentinula edodes

MES 02089 4B Su Xiang China Tested previously 1

MES 02008 sh 02/07 China Tested previously 2

MES 02110 le 03/13 Unknown Tested previously, commercial strain? 3

MES 02052 sh 02/02 China Tested previously 4

MES 02090 S600 Unknown Tested previously, commercial strain? 5

MES 00001 donko type 3715 Unknown Commercial strain 6

MES 00052 Lentinula edodes -2 Indonesia 7

MES 00061 Lentinula edodes -3 Indonesia 8

MES 02015 sh 03/01 Indonesia 9

MES 02016 sh 03/02 Indonesia 10

Pleurotus ostreatus

MES 14029 Florida Nigeria 11

MES 01997 pc 8-2-02 p2 Brasilia 12

MES 01998 pp 8-3-02 Brasilia 13

Pleurotus pulmonarius

MES 14030 Lagos Strain Nigeria 14

MES 14031 Ibadan Strain Nigeria 15

MES 11698 Jamur Amis - 3 Indonesia 16

MES 03448 pl 00/11 Unknown Commercial strain 17

MES 03444 G21 Unknown Commercial strain 18

Pleurotus eryngii

MES 12060 Le Lion PE Unknown Commercial strain 19

MES 03467 Sylvan 3065 Unknown Commercial strain 20

MES 02073 Mycelia 2600 Unknown Commercial strain 21

MES 03757 Pl02/08 China 22

MES 02001 Pl03/01 China 23

MES 12920 AL10 Iran 24

MES 12936 F21 Iran 25

MES 12961 M07 Kourdistan 26

MES 12970 M16 Kourdistan 27

MES 12929 D29 Iran 28

MES 12939 G02 Iran 29

MES 12978 S07 Kourdistan 30

Table 1. Species/strains used to test vegetative growth on cassava waste.

IV

were used to inoculate a single glass tube. After inoculation the strains were incubated at

24oC and checked at 3-5 day intervals for linear growth. Growth fronts were marked on 2

perpendicular axes on the side of the tube. For each strain linear growth was tested in 3

tubes (so called “race-tubes”). At the end of the experiment, the tubes were used for

determination of rumen digestibility. For this one tube has been frozen en freeze-dried after

2 weeks of colonisation, the second tube after 4 weeks and the last tube after 6 weeks.

Digestibility has been assessed after the method described by Cone et al. (1996). In short:

Rumen fluid was collected in the morning from two non-lactating Holstein–Friesian cows fed

1 kg of concentrate daily and grass silage ad libitum. Samples were incubated in triplicate

with rumen fluid for 72 h to determine gas production characteristics of the control and the

fermented substrates.

Mushroom cultivation trials on cassava waste

Peels and woody parts (sticks) of commercial cassava cultivation were send by FRI (Accra).

The woody parts comprised of sticks with a diameter of about 2-3 cm. The peels were sent

as dried material. The cassava waste materials were used to prepare 5 different substrates

(Table 2), three of which consisted solely of cassava waste material. Two consisted of

cassava waste materials mixed with saw dust (oak). As a control we included commercially

prepared wheat straw based substrate that is used by Dutch oyster mushroom growers.

Table 2. Substrate composition for small trials of mushroom production

Cassava stalks Cassava peels Saw dust CaCO3

Substrate 1 25% 75% 0% 1%

Substrate 2 50% 50% 0% 1%

Substrate 3 75% 25% 0% 1%

Substrate 4 25% 25% 50% 1%

Substrate 5 12.5% 12.5% 75% 1%

Substrate 6 Commercially available wheat straw based substrate (used by Dutch oyster

mushroom growers.

For the experiment 4 strains of Oyster mushroom were selected; Pleurotus pulmonarius

(lung oyster) strains MES03444 (commercial strains, originating from Slovakia) and

MES03448 (commercial strain, country of origin unknown) and Pleurotus ostreatus (grey

oyster) strains MES01997 and MES01998 (originating from Brazil). These strains were

chosen since they fructify at a higher temperature than strains used by European growers

(cultivation temperatures between 16 and 18 oC). Cassava peels are rich in starchy

components. In mushroom cultivation the presence of starchy components in substrates

poses a risk for contamination of the substrate with green moulds such a Trichoderma and

Aspergillus species. To reduce this risk, we subjected substrates 1 till 5 to fermentation at

elevated temperatures before inoculating the mushroom strains into the substrate.

Fermentation was performed in aerated boxes in an acclimatised room at an air

temperature of 43-45oC for a period of about 11 days. If substrates get warmed up to this

V

temperature, usually a thermophilic population starts to grow which heats up the substrate

to even higher temperatures. In our experiment the substrates only slightly elevated

temperatures were recorded. This indicates that the thermophilic population had difficulty

to grow. Likely, the low nitrogen content of this type of waste materials are the main cause

of low activities. Nevertheless, an extended period of elevated temperature will probably

kill many contaminating spores of fungal moulds. In households and small enterprises this

fermentation step can be done by “composting” waste on piles (or dikes) and leave it for 1

to 2 weeks with mixing the material every 2 to 4 days.

After fermentation, substrates were cooled to 24oC and inoculated with spawn of the Oyster

mushroom strains in a clean room. Strains were grown in plastic bags containing 2 kg of

inoculated substrate. After a 14 day period of spawn-run at 24oC, substrates were fully

colonised by the fungi. No infections with green moulds occurred. Mushroom production

was initiated by venting the room and lowering air temperature to 20oC. First mushrooms

started to appear 7 days after the onset of venting. Mushrooms were sampled for analysis

of dry weight, heavy metal content, presence of food pathogenic bacteria, mycotoxins,

pesticide residues and cyanide. These analyses are currently being performed.

6. Results

6.1 Tests on vegetative growth of fungal species/strains on cassava

waste.

The data are not complete since some strains need a very long time to colonize the

substrate and some still have not reached full colonisation after 44 days of colonization

time. These strains were excluded. Those strains that had reached full colonization of the

substrates are reported here and from these a selection was made for mushrooms growing

and upgrading waste into animal food. Colonization rate of substrate by a fungal strain is

well reflected by the linear growth rate of the strain on that substrate (figure 1).

Figure 2 shows the growth curve of L. edodes strain MES00052 on a mixture of cassava peels

and cassava stalks. In the period from 0 to 6 days the growth rate is slowly increasing. This

could be the adaptation phase in which the fungus needs to switch from feeding on malt

extract agar (in the inoculum points) to the plant material. However, this “lag” period can

also be due to a relatively poor contact between the agar plugs used for inoculation and the

Figure 1. Glass tubes filled with moistured and sterilized cassava waste and inoculated at the top with spawn of a fungal strain. The position of the growth front was measured every 3 days to estimate the colonization rate of each strain on different type of cassava wastes.

VI

rather uneven layer of plant material. After 6 days a long period with a nearly constant

growth rate can be seen. At the end of the incubation period, the growth rate seems to

diminish as the mycelium reaches the bottom of the tube. This may look like a depletion of

nutrients. However, it is more likely that during the long period of incubation some water

may have collected on the bottom of the tube. During the wetting of the substrate, excess

water (i.e. not taken up by the substrate and present in between the substrate particles) has

been pressed from the substrate as good as possible. However, small amounts of this excess

water may still have been present in the substrate when filling the tubes. After a long period

this water may have accumulated on the bottom of the tube. Such “free” water in the

substrate is known to retard fungal growth.

As the growth rates at the beginning and the end of the incubation period may not reflect

the true potential of the fungal strains, we have used only the linear part of the growth

curve for calculation of the growth rate (like is shown in Figure 3).

Results on growth of L. edodes strains on cassava based substrates are shown in Figure 4.

As can be seen for 7 out of 10 strains grow on all cassava waste types and the best growth is

shown on a 1:1 mixture of peels and sticks. Growth rate on a mixture varies from 2.6 to 3.5

mm/day at 24 oC.

Results on growth of P. ostreatus strains on cassava based substrates are shown in Figure 5.

The best growth of this species was on cassava sticks and growth rate varied between 6.5

and 7.1 mm/day. Growth rates on a mixture of peels and sticks was ca. 6.6 mm/day,

considerably faster than the L. edodes strains. For each combination of strain and substrate,

three replicates are present in the experiment. However, currently we do not have the data

for all replicates.

Figure 2. Growth curve of L. edodes strain MES00052 on a mixture of cassava peels and stalks.

Figure 3. Part of the growth curve used

for calculation of the linear growth rate.

VII

Results on growth of P. pulmonarius strains on cassava based substrates are shown in Figure

6. For this species growth is slowest on cassava peels, intermediate on a mixture of peels

and stalks and highest on cassava stalks, similar to growth rates of P. ostreatus. Growth of P.

eryngii strains on cassava based substrates are shown in Figure 7. Growth on cassava peels

was poor and only for 4 strains growth was recorded. Also this species showed a better

growth on sticks than on a mixture of sticks with peels.

Statistical analyses of the measurements are given in appendix I

Figure 5. Linear growth rate of P. ostreatus strains on cassava based substrates. As these are merely preliminary results, we can only show growth rate on a substrate consisting of a mix of cassava peels and stalks and a substrate based on only cassava stalks. Results on cassava peels only are not available yet. MES 14029 is a FIIRO strain.

0

1

2

3

4

5

6

7

8

MES 01997 MES 01998 MES 14029

P. ostreatus

Max

imu

m li

ne

ar g

row

th r

ate

(m

m/d

ay)

Cassava peels Cassava peels with 50% (w/w) stalks Cassava stalks

Substrate

Species Strain

Average of Linear growth rate (mm/day)

Figure 4. Linear growth rate of L. edodes strains on cassava based substrates. Growth on cassava sticks was very slow and discarded. Growth of strains that were very slow on peels and mixture of peels and sticks were also excluded.

0

0.5

1

1.5

2

2.5

3

3.5

MES 00001 MES 00052 MES 00061 MES 02008 MES 02015 MES 02016 MES 02052 MES 02089 MES 02090 MES 02110

L. edodes

Linear growth rate of Lentinula edodes (shiitake) on different cassava waste products


Substrate

Species Strain


VIII

Figure 6. Linear growth rate of P. pulmonarius strains on cassava based substrates. MES 14030 is a FIIRO strain.

0

1

2

3

4

5

6

7

8

MES 03444 MES 03448 MES 11698 MES 14030 MES 14031

P. pulmonarius

Max

imu

m li

ne

ar g

row

th r

ate

(m

m/d

ay)


Substrate

Species Strain


Figure 7. Linear growth rate of P. eryngii strains on cassava based substrates. As

these are merely preliminary results, we can only show growth rate on substrates consisting of a mix of cassava peels and stalks and a substrate based on cassava stalks only. Results on cassava peels only are largely not available yet.

0

1

2

3

4

5

6

MES 02001 MES 02073 MES 03467 MES 03757 MES 12060 MES 12920 MES 12929 MES 12936 MES 12939 MES 12961 MES 12970 MES 12978

P. eryngii

Max

imu

m li

ne

ar g

row

th r

ate

(m

m/d

ay)


Substrate

Species Strain


IX

6.2 Effects of fungal growth on digestibility by rumen microflora

After the completion of the vegetative growth tests, a selection of the tubes were used to

test the digestibility by rumen microflora as described in Cone et al., 1996. Gas production

was compared to non-inoculated (autoclaved) cassava waste. No effects was expected for

cassava peels since this material contains much starch. This starch will be consumed by

fungi before they will degrade the more fibrous material. The digestibility was, therefore,

tested with strains of one species only, i.e. P. pulmonarius. The digestibility of untreated

peels was considerably since the presence of starch (figure 8). As expected, the gas

production and thus the digestibility decreased after incubation with P. pulmonarius strains.

For the mixture of peels and sticks (1:1) an increase in gas production was seen with some

strains, especially those from L. edodes. The highest effect, however, was seen with the

cassava sticks. Sticks produce only minor amounts of gas in the rumen model as expected.

After especially treatment with strains from P. eryngii the amount of gas increased

considerably, up to levels close to 65% of peels showing potentials of these fungi to improve

digestibility of woody materials.

0.0

50.0

100.0

150.0

200.0

250.0

300.0

0 0 0 2 2 4 6 6

MES 11698 MES 03444 MES 11698 MES 11698 MES 03444

Uninoculated autoclaved control P. pulmonarius P. pulmonarius P. pulmonarius P. pulmonarius P. pulmonarius

Cassava Peels

Figure 8. Gas production (ml/gram organic matter) in the rumen model by cassava peels with or

without pretreatment with strains of P. pulmonarius. The digits under each bar indicate the

number of weeks of fungal pretreatment.

Figure 9. Gas production in a rumen model of different cassava waste products after fungal

pretreatment. The first panel shows the gas production by peels after treatment with P.

ostreatus. Gas production decreases compared to untreated samples probably due to

consumption of starch by the fungus. The second graph shows the gas production of a mixture

of cassava peels and stalks (1:1) after treatment with 2 Pleurotus species. Numbers under the

bars indicate the number of weeks of incubation. The third graph in figure 9 shows the gas

production of stalks after petreatment with the 3 Pleurotus species. Here, a clear increase of gas

production is seen compared to the untreated samples.

X

Figure 9.

XI

6.3 Mushroom production of selected strains on cassava waste

Two P. ostreatus and 2 P. pulmonarius strains were used to test mushroom yield on cassava

waste basted substrates. Mushrooms were harvested during a period of ca. one month (2

flushes for each strain). The production of the strain used by Dutch growers (SPOPPO) on

commercial substrate (wheat straw) did produced earlier (6 days after climate change to

induce fruiting) than all other treatments. The best production was seen by the commercial

strain used by Dutch growers on commercially available substrate (wheat straw). This is not

unexpected since the strain/substrate has been developed especially for cultivation in Dutch

systems (similar to what we have used here). Also, Dutch substrate is supplemented with

nitrogen and the cassava waste based substrates has been used either as they are or

supplemented with oak tree saw dust (low in nitrogen). Despite the absence of supplements

it is remarkable that yields on cassava waste only are reasonably good, varying from 6% up

to more than 15% (figure 8). The highest yield is seen on a 50:50 mixture of cassava peels

and sticks with a Brazilian P. ostreatus strain. If yields are compared for cassava based

substrates only (with or without oak saw dust) and averaged for all strains, a mixture of

peels and stalks (1:1) shows the best production (figure 9). It is surprising that the addition

of saw dust had no positive effect on the production. This might be either due to the low

quality of the saw dust or the inability of the fungi to utilize saw dust well without enough

nitrogen available.

Figure 8. Yield of oyster mushroom strains of different substrates expressed as Biological

Efficiency (fresh weight mushrooms/fresh weight substrate). The commercial wheat straw

substrate (substrate 6) has the highest yields and is supplemented with nitrogen, contrary to the

other cassava waste based substrates. Nevertheless, yields up to 15% can be reached.

XII

Substrate bags were weighted at the beginning and at the end of the cultivation. In this way

yields of mushrooms can be compared with the decrease in weight of the substrate. A very

clear correlation was seen (figure 10) indicating the good quality of the data.

0

50

100

150

200

250

300

Substrate 1 Substrate 2 Substrate 3 Substrate 4 Substrate 5

Gra

mm

es

mu

shro

om

s p

er

2 k

g su

bst

rate

Total yield in 2 flushe

substraat

Average of Totale opbrengst 2 vluchten (g)

Figure 9. Yield (gram mushrooms per bag=2 kg substrate) for substrates based on

cassava waste only. The mixture of peels and stalks gave the best production.

Figure 10. Correlation between mushroom yield and decrease of

substrate weight. A clear correlation is shown indicating that the

measurements were sound.

y = 0.0014x + 0.0218R² = 0.8329

0

0.2

0.4

0.6

0.8

1

1.2

0 100 200 300 400 500 600 700 800De

cre

ase

su

bst

rate

we

igh

t (k

g)

Yield mushrooms (grammes)

Correlation between yield and decrease substrate weight

XIII

7. Conclusions

The main conclusions of this research are:

At least 4 different fungal species used to produce edible fungi grow well

vegetatively on cassava waste (peels, stalks and a mixture of both (1:1)). Comparison

of growth rate for the 4 species tested

o L. edodes: Mixture of peels & stalks> peels or stalks alone

o P. ostreatus: Stalks> mixture> peels

o P. pulmonarius: Stalks> mixture> peels

o P. eryngii: Stalks> mixture> peels

Pretreatment of especially stalks with P. eryngii increases the digestibility for rumen

flora considerably.

The production of grey oyster mushrooms (P. ostreatus) on substrate consisting of

cassava waste only is good (up to 15% fresh weight mushrooms/fresh weight

substrate).

Experiments are started now to test yields on cassava waste supplemented with

nitrogen rich waste. We expects that addition (like bran) can increase yields up to

levels of commercial production.

XIV

8. References

Cone, J.W., van Gelder, A.H., Visscher, G.J.W., Oudshoorn, L., 1996. Influence of rumen fluid

and substrate concentration on fermentation kinetics measured with a fully automated time

related gas production apparatus. Anim. Feed Sci. Technol. 61, 113–128.

Aro S O, Aletor V A, Tewe O O and Agbede J O 2010: Nutritional potentials of cassava tuber

wastes: A case study of a cassava starch processing factory in south-western Nigeria.

Livestock Research for Rural Development. Volume 22, Article #213. Retrieved December 6,

2013, from http://www.lrrd.org/lrrd22/11/aro22213.htm

Annes 1 Pictures of mushroom production

Annex 2 Statistical analysis of yield data

Program used; Genstat, 16th edition.

Raw data

Strain Substrate Tray Yield (g) Strain Substrate Tray Yield (g)

MES 01997 Substrate 1 25 247 MES 03444 Substrate 1 6 193


XV


























XVI






















Analysis of variance.

Variate: Yield_g

Source of d.f. s.s. m.s. v.r. F pr.

XVII

variation

Tray stratum

Strain 3 390814 130271 47.98 <.001

Substrate 5 1342896 268579 98.92 <.001

Strain.Substrate 15 43355 2890 1.06 0.404

Residual 72 195492 2715

Total 95 1972558

Information summary

All terms orthogonal, none aliased.

Message: the following units have large residuals.

Tray 82 111.8 s.e. 45.1

Tray 95 -141.8 s.e. 45.1

Tables of means

Variate: Yield_g

Grand mean 260.1

Strain MES 01997 MES 01998 MES 03444 MES 03448

331.4 223.0 175.1 311.1

Substrate Substrate

1

Substrate

2

Substrate

3

Substrate

4

Substrate

5

Substrate

6

228.9 242.4 213.5 200.4 157.8 517.9

Substrate

Strain Substrate Substrate Substrate Substrate Substrate Substrate

XVIII

1 2 3 4 5 6

MES

01997

265.0 309.7 310.0 282.0 249.5 572.0

MES

01998

208.0 204.5 150.0 164.2 131.5 479.5

MES

03444

178.2 132.5 136.2 122.0 74.5 407.2

MES

03448

264.2 322.8 257.8 233.5 175.8 612.8

Standard errors of differences of means

Table Strain Substrate Strain x Substrate

rep 24 16 4

d.f. 72 72 72

s.e.d. 15.04 18.42 36.85

Least significant differences of means (5% level)

Table Strain Substrate Strain x Substrate

rep 24 16 4

d.f. 72 72 72

l.s.d. 29.99 36.73 73.45

Strain Substrate

Average yield (g)/2 kg of

substrate

(wet weight/wet weight)

l.s.d. is 73.45 at P=0.05

XIX

MES 03444 Substrate 5 75 a

MES 03444 Substrate 4 122 a b

MES 01998 Substrate 5 132 a b c

MES 03444 Substrate 2 133 a b c

MES 03444 Substrate 3 136 a b c d

MES 01998 Substrate 3 150

b c d e


b c d e f


b c d e f


b c d e f g


c d e f g h


e f g h


f g h i


g h i j


h i j


h i j


h i j


i j


j


j


j


k


k


l


l

Values sharing the same letter show not statistically significant difference at p=0.05

XX

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DELIVERABLE REPORT - fp7-gratitude.eu · MES 11698 Jamur Amis ... MES 03444 G21 Unknown Commercial strain 18 Pleurotus eryngii MES 12060 Le Lion PE Unknown Commercial strain 19 MES