Acta Astronautica 63 (2008) 1037 1042 www.elsevier.

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Bioconversion of rice straw into a soil-like substrate

Chengying Yua , Hong Liub , , Yidong Xingc , N.S. Manukovskyd , V.S. Kovalevd , Yu.L. Gurevichd
a Department of Environmental Engineering, Beijing University of Aeronautics and Astronautics, Beijing 100083, China b Department of Bioengineering, Beijing University of Aeronautics and Astronautics, Beijing 100083, China c Department of Applied Chemistry, Beijing University of Aeronautics and Astronautics, Beijing 100083, China d Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Krasnoyarsk 660036, Russia

Received 27 June 2007; received in revised form 21 December 2007; accepted 23 March 2008 Available online 13 May 2008

Abstract To increase the closure of bioregenerative life support systems (BLSS), the bioconversion of rice straw into a soil-like substrate (SLS) by mushrooms and worms has been studied. The results showed that rice straw could be treated better by aerobic fermentation and succeeding growth of mushrooms Pleurotus ostreatus. In this process the total content of lignocellulose in the straw was removed by 37.74%. Furthermore, 46.68 g (fresh weight) of mushrooms could be produced from 100.0 g (dry weight) of rice straw. During the conversion of rice straw into a starting SLS by mushrooms and worms, the matter loss was 77.31%. The lettuce has been planted in the SLS and the yield when lettuce was cultivated on the SLS (8.77 g m2 day1 ) was comparable to the yield obtained on the nutrient solution. In addition, the silicon in the SLS ash can reach upto 32% and the circulation of it is expected during the growth of rice. 2008 Elsevier Ltd. All rights reserved.
Keywords: Life support systems; Soil-like substrate; Matter recycling; Rice straw; Silicon

1. Introduction Bioregenerative life support systems (BLSS) with highly closed intrasystem mass exchange hold much promise for long-term human life support in Lunar or Mars base, BLSS where higher plants act as a photosynthesizing component has been demonstrated to have good prospects. In order to develop a life support system with highly closed internal mass exchange, the efcient operation of the photosynthesizing component in such a system and maximum possible utilization of plant wastes (inedible for humans) are topical issues.
Corresponding author. Fax: +86 10 82339837.

E-mail address: lh64@buaa.edu.cn (H. Liu). 0094-5765/$ - see front matter 2008 Elsevier Ltd. All rights reserved. doi:10.1016/j.actaastro.2008.03.010

About the utilization of plant wastes, many physicochemical and biological methods were studied, such as incineration technology, aerobic microbial decomposition and so on [1,2]. The Institute of Biophysics in Russia (Russian Academy of Sciences, Siberian Branch) has developed the process of producing a soil-like substrate (SLS) from plant wastes by mushroom and worms, and the SLS could be used as a substrate for growing plants [3]. It was shown that wheat, bean, cucumber and radish could grow on the SLS, and their yields when cultivated on a mature SLS were comparable to the yields obtained on a neutral substrate (expanded clay aggregate) under hydroponics [4]. The dynamics of residual mass conversion and the time when the system with an SLS reached steady state have been evaluated [5,6].

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Table 1 The growing conditions of mushrooms
Phase Temperature ( C ) 22 23 13 Humidity (%) 60 90 90 Illumination ( mol m2 s1 ) 0 15 0

A possible BLSS layout where SLS is used to grow higher plants and utilize inedible phytomass has been considered [3]. Two kinds of plant wastes (wheat straw and inedible potato biomass) have been tested to convert into SLS [7]. The distinctive feature of rice straw is that its silicon content (4.53%) is higher than that in wheat straw (1.28%) and it may be much more difcult to process the rice straw [8]. The purpose of this work is to test a two-stage biological conversion of inedible rice straw into an SLS by means of oyster mushrooms and worms and to nd out an appropriate processing method and an optimal mushroom. The leaf lettuce was selected in the list of candidate plants in BLSS for a lunar base [9], and it was also used as a model salad crop because of its short production cycle, its adaptability to controlledenvironment cultivation and its relatively uncomplicated growth pattern [10]. Hence, the leaf lettuce was used in the SLS testing. 2. Materials and methods 2.1. Wastes material The initial material for the SLS was the eld straw from rice plants with added rice bran. The mass ratio of bran to rice straw was 10%. The rice straw and bran were supplied by agricultural experimental base in Beijing suburb. 2.2. Organisms The cultures of mushrooms (Pleurotus ostreatus, Pleurotus sapidus, Pleurotus sajor-caju and Pleurotus orida) and worms (Eisenia foetida Savignnny) were maintained at the Laboratory of Environmental Engineering Department of Beijing University of Aeronautics and Astronautics. The lettuce (Lactuca sativa var. capatata L.) seed was provided by The Chinese Academy of Agricultural Sciences. 2.3. Preparation of the substrates from the wastes The substrate with 65% water consisted only of rice straw, which was cut rstly (fragment size 13 cm) and bran. Half of the substrate was placed into vessels and sterilized for 2 h in steam at a pressure of 11.2 atm in a sterilizer. After sterilized and cooled, the substrate in the jars was prepared to inoculate with oyster mushroom. The other half of the straw without being inoculated with any micro-organisms was fermented at 60 C for 24 h and then at 45 C for two days in the facility. After

Spawn running Fruiting Day Night

fermented and cooled, the straw was placed in vessels. The cultivation vessels were 32 plastic bags. Each vessel contained 500 g of mixed substrates in terms of wet weight. 2.4. Substrate conversion by oyster mushroom The substrate in the bags was inoculated with oyster mushroom spawn grown on the wheat grains under sterile conditions. The spawn was added to the straw in a mass ratio of 1 part spawn per 10 parts straw. All the mushrooms used in the experiment are edible and bre degraders [11]. After inoculation, the bags were placed in the growth chamber. The sterile and other conditions were kept under spawn running. When spawn running is nished, the mushrooms initiated fruiting and some conditions such as humidity and illumination were changed (Table 1). The efciency of waste processed by mushrooms was estimated by the mushroom fruit bodies production, loss of matter and degradation of lignocellulose. The fruit production was evaluated by the biological efciency calculated as the ratio of wet mass of fruit bodies to the initial dry mass of the substrate in the cultivation bag. The loss of matter was calculated as the ratio of initial masses. 2.5. Residual substrate conversion by worms When mushroom growth was completed, the spent mushroom compost was transferred from plastic bags to boxes with 35 cm 45 cm 30 cm in dimensions and converted to the starting SLS by worms. The density of adult worms was 20 5 worms per 100 g of wet substrate mass at rst and the worms were cultivated for four months. During this period, the water content of the substrate was maintained at 7580%, and the temperature was maintained at 2025 C. The starting SLS was a mixture of residual substrate and worms, and it was tested by the lettuce growing on it.

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2.6. Testing SLS by plant growth The lettuce was planted in plastic basins (17 cm diameter on the top and 12 cm at the bottom by 14 cm deep). The substrate layer was about 10 cm thick. Each box contained about 0.7 kg of SLS in terms of wet mass or 0.21 kg of dry mass. The farmland soil was used as the comparison and its weight was about 2 kg of wet mass or 0.6 kg of dry mass. The lettuce seedlings were cultivated in soil substrate previously. When having 45 leaves, the seedlings were transplanted into the SLS at a density of about 54 plants per 1 m2 (1 plant per basin). The lettuce growing on the SLS was watered once or twice every two days by irrigation water and the water content of the SLS and farmland soil was maintained at 70%. The lettuce was grown at a temperature of 2023 C and a continuous illumination of 250300 mol m2 s1 (photosynthetic photon ux). The air moisture content was maintained at 70%. After 4050 days, the lettuce was harvested and the total and edible masses of lettuce were determined. The fertility of SLS and farmland soil was indicated by the yields of total (Yt , Yt ) and edible (Ye , Ye ) lettuce biomass. The values of Yt , Yt , Ye and Ye were calculated using the following formulae: Yt = Mt /S ; Yt = Mt /Ms ; Ye = Me /S ; Ye = Me /Ms , (1) (2) (3) (4)

3. Results and discussion 3.1. Substrate pretreatment comparison and mushrooms selection In the experiment, the results proved that the mushroom spawn can convert and utilize the nutrition of rice straw with high silicon content and fruit. After sterilization by autoclave or aerobic fermentation, the straw substrate was used to culture mushroom, and the biological efciency, lignocellulose removal ratio and the yield of mushrooms are presented in Table 2. As the results indicated, for the four mushrooms, the fermented straw was converted better and produced more mushrooms than the straw sterilized by autoclave more or less. During the process, sterilization by autoclave can be nished just in 23 h, while it must use airtight vessels resisting the high pressure. What is more, the substrate should be encased into the bags before sterilization. In this case, the substrate can be inoculated only on the top with a mushroom spawn. While after fermentation, the substrate can be encased into the bags when inoculating, and the spawn can be mixed with a substrate evenly on the large contacting area and grow faster. However, the disadvantage of fermentation is its long period; it needs about 36 h to nish. Furthermore, the fermented substrate has a comparative advantage to prevent infection during inoculation. Some benecial microorganisms might get the advantage and restrain some harmful microorganisms. The probability of pollution will be reduced. To sum up, the aerobic fermentation is a better process for the pretreatment of straw. Four varieties of mushrooms were compared by the degradation of rice straw, and the yield and growth period of mushrooms were considered too. The indexes of the mushrooms, after they were grown in the fermented substrate, are given in Table 2. The P. ostreatus mushroom showed preferable character. In this process, 46.68 g (fresh weightFW) of mushrooms could be produced from 100.0 g (dry weightDW) of rice straw, and the biological efciency and yield were 46.68% and 10.15 g kg1 day1 , respectively. Furthermore, the lignocellulose in straw was removed by 37.74%. The loss of matter was 48%, which was lower than 78% in the wheat straw conversion [7]. It is because the mushrooms were harvested only once after 4050 days growth, while the period of mushrooms that grew on the wheat straw substrate was 6070 days. But the extension of mushroom growth will undoubtedly prolong the total period and the treatment efciency of straw

where Mt is the total dry (or fresh) biomass of lettuce per cultivation vessel upon completion of its growth, Me is the dry (or fresh) edible biomass of lettuce per cultivation vessel upon completion of its growth, Ms is the dry (or wet) mass of the substrate and S is the cultivation vessel area.

2.7. Chemical analysis The fresh and dry masses, lignocellulose, ash and silicon contents in the samples were determined by standard methods [12]. The silicon content was calculated from the silicon dioxide in the samples. The available major and trace elements of the samples were analysed by ASI means [13].

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Table 2 The conversion of rice straw by mushrooms after different pretreatments

Mushrooms Biological efciency (%) Yield of mushrooms (FM/DMa ) (g kg1 day1 ) Lignocellulose removal ratio (%) Matter loss (%)
a FM/DMfresh b Athe

P. sapidus Ab Fc A F A F A F 32.69 36.17 6.954 6.562 22.17 29.05 24.42 39.73

P. sajor-caju 23.41 31.65 3.657 8.554 21.45 23.15 22.56 34.60

P. ostreatus 33.77 46.68 7.342 10.15 24.80 37.74 23.72 48.02

P. orida 21.28 23.40 5.190 5.707 22.12 24.98 20.33 29.86

mass of mushrooms per dry mass of the substrate. rice straw substrate after sterilization by autoclave. c Fthe rice straw substrate after aerobic fermentation.

Table 3 Some available components of SLS (g kg1 )

Elements Values NO 3 N 3.184 NH+ 4 N 0.221 P 1.639 K 16.480 Cu 0.008 Fe 0.058 Mn 0.292 Zn 0.047 Ca 5.303 Mg 3.660 S 0.987 B 0.015

per unit time will reduce relatively. Our calculation indicated that the yield of mushrooms per unit time from rice straw10.15 g kg1 day1 was larger than that from wheat straw7.74 g kg1 day1 [4]. 3.2. Converting spent mushroom compost to the starting SLS The spent mushroom compost was processed by worms followed. After four months, the substrate softened and changed from yellow to dark brown. This articial soil can be called an SLS. In the conversion of rice straw to the starting SLS, the matter loss was 77.31%. It could be associated with the production of carbon dioxide and metabolic water. Some available components of SLS are presented (Table 3) and we may draw a conclusion that the nutrition of the SLS converted from rice straw by microorganisms, mushrooms and worms can be supplied to plants directly in theory. 3.3. Testing SLS fertility The yields of lettuce cultivated on the SLS and farmland soil are shown in Fig. 1. The calculations showed that the yield of the edible part of lettuce can reach upto 6.8 g kg1 day1 (FW/DW), which is larger by 30% than that on the farmland soil. The yield of the edible part of lettuce calculated per unit area was 8.77 g m2 day1 (DW), which was comparable to 7.10 g m2 day1 dry biomass harvested from

a modied strength Hoagland nutrient solution (16-h light-300 mol m2 s1 /8-h dark photoperiod) [14]. However, the yield calculated per unit area on the SLS was lower than that on the farmland soil (18.69 g m2 day1 ). The difference may be accounted for by the fact that there is much more weight of the soil in the same vessel volume. The result indicates that many problems about the SLS should be studied henceforth. The thickness of the SLS, plant density and illumination were not the best for lettuce growth in the experiment, and the yield may increase if these indexes can be optimized. To sum up, it is feasible that the SLS is used to plant lettuce and produce some biomass. The components of lettuce harvested on the SLS converted from rice straw were not tested, but the plants such as wheat, beans, radishes and cucumbers gained from the wheat straw SLS have been tested. The testing demonstrated that there were no signicant differences in the majority of the biochemical characteristics used to evaluate the edible part of plants grown under hydroponics [15] and on the SLS [6,16,17]. 3.4. Silicon in SLS The high silicon content is a problem that researchers have been paying attention to in the treatment of rice straw. The changes in silicon and ash contents in samples at different treatment phases of rice straws are shown in Fig. 2. The ash content change was approximately uniform with silicon shown in the gure, and

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60
10 9 8 yield (gkg-1 day-1) 7 6 5 4 3 2 1 0 TM(F/W) TM(F/D) EM(F/W) EM(F/D) SLS farml and soil

50 Proportion (%) 40 30 20 10 0 1 2

Si ash

200 180 160 yield (gm-2 day-1) 140 120 100 80 60 40 20 0 TM(F) TM(D) EM(F) EM(D) SLS farml and soil

3 Steps

Fig. 2. The changes in silicon and ash contents in the samples at different treatment steps. 1rice straw; 2rice straw after fermentation; 3rice straw after mushroom growing; 4SLS; 5SLS after lettuce growing.

ing. If rice is introduced into the plantsSLS system, it is expected that silicon can be circulated unceasingly as other elements, which complies with the initial idea of the SLS design.

Fig. 1. The yields of lettuce harvested on the SLS and farmland soil. (a) Yield of lettuce calculated per unit mass of substrate; (b) yield of lettuce calculated per unit area. TM (F/W)ratio of total fresh mass of lettuce to wet mass of the substrate; TM (F/D)ratio of total fresh mass of lettuce to dry mass of the substrate; EM (F/W)ratio of edible fresh mass of lettuce to wet mass of the substrate; EM (F/D)ratio of edible fresh mass of lettuce to dry mass of the substrate; TM (F)total fresh mass of lettuce; TM (D)total dry mass of lettuce; EM (F)edible fresh mass of lettuce; EM (D)edible dry mass of lettuce.

4. Conclusions The experiments carried out have shown that rice straw could be converted into an SLS by biological (microorganisms, mushrooms and worms) processing. Rice straw could be treated better by aerobic fermentation and succeeding growth of the P. ostreatus mushrooms. In the conversion of rice straw to an SLS, the matter loss was 77.31%. The starting SLS containing high nutrition was gained, and 8.77 g m2 day1 (DW) edible part of lettuce was harvested on it. The experiments have shown that SLS converted from rice straw could be used to grow crops in BLSS. Further research is planned to focus on evaluating the characteristics of multispecic plant communities grown on the SLS. It is necessary to study the organic matter turnover and the transformation of silicon in rice straw SLS during planting, and the growing indexes of candidate plants should be optimized.

the silicon content was nearly 32% of the ash in the nally obtained SLS. Compared with 19% ash in the wheat straw SLS [7], the ash content in the SLS converted from rice straw is 45% and the main difference is caused by the silicon content. After lettuce growing, the contents of silicon and ash continued rising, because the organic matter in the SLS was decomposed and used under the combined actions of lettuce, worms and microbes. However, silicon and ash were hardly absorbed by crops; therefore, they cumulated in the SLS. At the beginning of the experiment, the absorption of silicon is little if only taking vegetables for the experiment, and there will be accumulation of silicon in the SLS after a period of time. However, it is well known that rice absorbs much silicon when grow-

Acknowledgements This research was sponsored by the Ministry of Science and Technology of China Grant 2005DFBA005.

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