AU2021102640A4 - An apparatus for sugarcane propagation and a method thereof - Google Patents
An apparatus for sugarcane propagation and a method thereof Download PDFInfo
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- AU2021102640A4 AU2021102640A4 AU2021102640A AU2021102640A AU2021102640A4 AU 2021102640 A4 AU2021102640 A4 AU 2021102640A4 AU 2021102640 A AU2021102640 A AU 2021102640A AU 2021102640 A AU2021102640 A AU 2021102640A AU 2021102640 A4 AU2021102640 A4 AU 2021102640A4
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/55—Sugar cane
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
- A01H6/46—Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
An apparatus for sugarcane propagation, the apparatus comprises of a
basal medium 100 for regenerating roots of sugarcane, wherein the
basal medium 100 comprising of a sterilized glassware containing a
defined quantity of double distilled water and a plurality of chemicals
and stock solutions in required amount, at least a container 102
autoclaved at 110-130 degree Celsius for 15-25 minutes and stored
under light for about 15-20 hours and under dark for about 7-9 hours
for inducing shoot and forming roots, at least a portray for
accommodating at least a heterotrophic micro-propagated plant
containing a mixture of soil, sand and cocopeat in a defined ratio, and
a hardening cage for culturing the heterotrophic micro-propagated
plants into an autotrophic and acclimatized sturdy plant, wherein the
hardening cage comprises of a shade net for allowing light to reach the
plants for growth.
24
100 106
BASAL MEDIUM HARDENINGCAGE
102 108
CONTAINER THERMOMETER
110
104
PROTRAY HYGROMETER
112
LUXMETER
FIGURE 1
regenerating roots of sugarcane in a basal medium, wherein the steps of regeneration comprising of:
202
a) discloses about adding a defined quantity of double distilled water, a plurality of chemicals and
stock solutions in required amount into a sterilized glassware 202a
b) autoclaving at least a container at 110-130 degree Celsius for 15-25 minutes and storing under
light for about 15-20 hours and under dark for about 7-9 hours for inducing shoot and forming roots.
202b
c) accommodating at least a heterotrophic micro-propagated plant in at least a portray, wherein the
pRotra contains a mixture of soil, sand and cocopeat in a defined ratio.
202c
culturing the heterotrophic micro-propagated plants into an autotrophic and acclimatized sturdy
plant inside a hardening cage, wherein the hardening cage comprises of a shade net for allowing light \
to reach the plants for growth 204
FIGURE 2
Description
106 BASAL MEDIUM HARDENINGCAGE
102 108 CONTAINER THERMOMETER 110 104 PROTRAY HYGROMETER
112 LUXMETER
FIGURE 1
regenerating roots of sugarcane in a basal medium, wherein the steps of regeneration comprising of:
202
a) discloses about adding a defined quantity of double distilled water, a plurality of chemicals and stock solutions in required amount into a sterilized glassware 202a
b) autoclaving at least a container at 110-130 degree Celsius for 15-25 minutes and storing under light for about 15-20 hours and under dark for about 7-9 hours for inducing shoot and forming roots. 202b c) accommodating at least a heterotrophic micro-propagated plant in at least a portray, wherein the pRotra contains a mixture of soil, sand and cocopeat in a defined ratio. 202c
culturing the heterotrophic micro-propagated plants into an autotrophic and acclimatized sturdy plant inside a hardening cage, wherein the hardening cage comprises of a shade net for allowing light \ to reach the plants for growth 204
FIGURE 2
The present invention generally relates to a field of propagation in plants. More particularly, the present invention relates to an apparatus and a method for micropropagation of sugarcane plant.
Micropropagation has been extensively used for the rapid multiplication of many plant species. However, the use of this method in sugarcane propagation is restricted by loss or damage of plants when transferred to ex vitro conditions like greenhouse or field. The overall success of tissue culture raised plants depends on successful hardening and transplantation in the field. After obtaining the in vitro raised plants, they are required to be carefully hardened and acclimatized. The readily available nutrient supply and controlled environmental conditions during in vitro cultures do not require any change in anatomical and morphological conditions in micro propagated plants.
However, plantlets which are cultured in vitro on agar-based media wilt rapidly on transfer to normal, green house or field conditions. Poor water uptake and rapid loss of water through transpiration may lead to high mortality rate unless plantlets are acclimatized by gradually exposing plantlets to reduced humidity and increased light intensity. The process of establishment of plantlets is compounded by damage to the delicate shoots and roots during transplantation. Moreover, good drainage and sufficient aeration of roots are essential factors for the luxuriant growth of the transferred plants.
Micropropagation of sugarcane is achieved through the establishment of explants, their initial growth in vitro followed by transplanting in greenhouse or field. During in vitro culture, plantlets grow under relatively high air humidity with low irradiance in closed vessels in order to prevent microbial contamination and to decrease air turbulence which prevent inflowof C02 and outflow of gaseous plant products from the vessels.
The final stage of in vitro clonal propagation involves hardening and establishment of plantlets to the ex vitro environment in soil. Inevitably, tissue culture conditions which promote rapid growth and proliferation result in formation of abnormal plants with altered leaf morphology, mesophyll structure, poor photosynthetic activity, malfunctioning of stomata and reduced cuticular waxes. During transfer of plants to soil and hardening, desiccation of shoots due to substantial loss of water through cuticular and stomatal transpiration and change from heterotrophic to autotrophic mode in nutrition are the major reasons for higher shoot mortality.
Therefore, methods for acclimatization of plants under greenhouse or field conditions need to be optimized to achieve higher rates of survival during temperature extremes.
Existing method leads to excessive water loss in the micro-propagated plants resulting in lower survival rates in the field.
In order to overcome the above-mentioned limitations, there exists a need to develop an apparatus to study the effect of the single shade net on hardening of selected sugarcane genotypes in terms of temperature, light intensity and humidity.
The technical advancements disclosed by the present invention overcomes the limitations and disadvantages of existing and convention systems and methods.
The present invention generally relates to an apparatus for propagation of sugarcane plant.
An objective of the present invention was to use sugarcane genotypes for micropropagation of sugarcane.
Another objective of the present invention was to provide a modified method of propagation in sugarcane.
Another objective of the present invention was to study the hardening of selected sugarcane genotypes in terms of temperature, light intensity and humidity.
According to an aspect of the present disclosure, the apparatus comprises of a basal medium in a sterilized glassware, a container, a portray, a hardening cage, a thermometer, a hygrometer and a lux meter.
The basal medium for regenerating roots of sugarcane, wherein the basal medium was provided in the sterilized glassware.
The Basal medium contain amino acids, glucose, and ions (calcium, magnesium, potassium, sodium, and phosphate) essential for cell survival and growth. I-glutamine is an amino acid that is essential for protein and nucleic acid synthesis and energy production in a cell culture.
The sterilized glassware containing a defined quantity of double distilled water and a plurality of chemicals and stock solutions in required amount.
Sterilization refers to any process that removes, kills, or deactivates all forms of life (in particular referring to microorganisms such as fungi, bacteria, spores, unicellular eukaryotic organisms such as Plasmodium, etc.) and other biological agents like prions present in a specific surface, object or fluid.
The glassware is preferably a bowl, a tumbler, a plate or a glass.
The container was autoclaved at 110-130 degree Celsius for 15-25 minutes and stored under light for about 15-20 hours and under dark for about 7-9 hours for inducing shoot and forming roots.
The term 'autoclave' herein refers to a machine that provides a physical method of sterilization by killing bacteria, viruses, and even spores present in the material put inside of the vessel using steam under pressure. Autoclave sterilizes the materials by heating them up to a particular temperature for a specific period of time.
The temperature and the duration of autoclaving differs according to the requirement of the apparatus.
The container includes a pre-sterilized culture bottle holding a liquid medium of 40-50ml, wherein the container is autoclaved at 121 degree Celsius at 15|bpsi for about 15-25 minutes.
The containers are placed under the light source at a height difference of 25-30cm, wherein subculturing is done after every two-three weeks.
The container is accommodated with well rooted with at least 16 genotypes, wherein the genotypes are obtained through an in-vitro culture of apical meristem from top shoot of each of the plurality of sugarcane stock plants.
The protray is for accommodating at least a heterotrophic micro propagated plant containing a mixture of soil, sand and cocopeat in a defined ratio, wherein the ratio of soil, sand and cocopeat in the portray is 1:1:1 respectively.
The hardening cage is used for culturing the heterotrophic micro propagated plants into an autotrophic and acclimatized sturdy plant, wherein the hardening cage comprises of a shade net for allowing light to reach the plants for growth.
The term hardening is a process where heterotrophic in vitro cultured delicate plantlets become autotrophic and acclimatized sturdy plants. This hardening process is carried out in the structures called hardening cage.
The hardening cage may be a single shade cage, a double shade cage or a polyhouse. The hardening cage used herein is a single shade cage. The shade net house is constructed with a green shade net permitting 50 per cent light and the average temperature, relative humidity and light intensity were 28.3 0C, 63.20 per cent and 7522 lux, respectively. The thermometer, the hygrometer and the lux meter are used to monitor temperature, relative humidity and light intensity respectively in all three hardening cage structures.
According to an aspect of the present disclosure, the method for sugarcane propagation, the method comprises of:
The first Step discloses about regenerating roots of sugarcane in a basal medium, wherein the steps of regeneration comprising of: a) adding a defined quantity of double distilled water, a plurality of chemicals and stock solutions in required amount into a sterilized glassware.
b) autoclaving at least a container at 110-130 degree Celsius for 15-25 minutes and storing under light for about 15-20 hours and under dark for about 7-9 hours for inducing shoot and forming roots.
c) accommodating at least a heterotrophic micro-propagated plant in at least a portray, wherein the protray contains a mixture of soil, sand and cocopeat in a defined ratio.
The second step discloses about culturing the heterotrophic micro propagated plants into an autotrophic and acclimatized sturdy plant inside a hardening cage, wherein the hardening cage comprises of a shade net for allowing light to reach the plants for growth.
According to an aspect of the present disclosure, the different hardening conditions are single shade net, double shade net and polyhouse.
The shade net house is constructed with green shade net permitting per cent light and the average temperature, relative humidity and light intensity were 28.3 0 C, 63.20 per cent and 7522 lux, respectively.
The double shade net hardening cage structure made with two layers of green shade net and the average temperature, relative humidity and light intensity were 31.39 0C, 68.88 per cent and 5210.3 lux, respectively.
The Polyhouse hardening structure constructed with UV stabilized poly sheet and the average temperature, relative humidity and light intensity were 38.03 0C, 78.14 per cent and 31685.7 lux, respectively.
The thermometer, hygrometer and lux meter are used to monitor temperature, relative humidity and light intensity respectively in all three hardening cage structures.
The characteristics considered for monitoring the growth are shoot length, number of leaves, chlorophyll content (mg/g) and root characteristics: number of roots, root length, root volume and survivability.
The response of a first genotype C85004 under different hardening cage conditions. For the first genotype Co 85004 single shade net showed better response for all the characteristics except chlorophyll content which recorded highest value (21.80 mg/g) in double shade net and better survivability ( 2 1.5 0 %) in polyhouse.
The response of second genotype Co0323 under different hardening cage conditions. Single shade net is also found optimum for hardening of Co 0323 considering all characteristics measured except mortality percentage (7.5 0 %) for which double shade net was the best.
The response of third genotype CoVC 07-06-05, fourth genotype CoVC 09-65-04, a fifth genotype CoVC 99463, a sixth genotype CoVC 09 61-07, and a seventh genotype CoVC 00964 under different hardening cage conditions. single shade net was found optimal for all shoot morphological characteristics.
The response of eighth genotype Co 99004 under different hardening cage conditions. The optimum response for hardening of Co 99004 is in single shade for all characters except shoot length (36.93) and number of leaves (3.33) for which polyhouse was found to be the best. The degree of adaptation is to be increased by growing shoots in reduced humidity before transfer, but shoot growth and rooting capacity may decline progressively as humidity is lowered.
The response of ninth genotype Co62175 under different hardening cage conditions. The single shade net showed better response for all characteristics except for the number of leaves (Table 5). For number of leaves the highest value is observed (3.07) in double shade net and better survivability recorded ( 3 2 .5 2 %) in polyhouse in the genotype Co 62175.
The response of tenth genotype CoVC 10-65-01 under different hardening conditions. Under single shade net, CoVC 10-65-01 showed better response for all characteristics except shoot length and for which highest (39.60) value is observed in double shade net with better survivability (7. 3 5%) recorded in polyhouse.
The response of eleventh genotype CoT 12368 under different hardening conditions. The genotype CoT 12368 showed better response under single shade net for the four characteristics chlorophyll content, root volume, number of roots and root length. For shoot length (40.27), number of leaves (3.13) and mortality percentage (10.1 7% ) polyhouse was found to be best.
The response of twelfth genotype CoVC 10-43-06 under different hardening conditions. Single shade net is found better for the genotype CoVC 10-43-06 considering chlorophyll content, root volume, number of roots and root length except shoot length (27.40) and mortality percentage (17. 48 %). For these two traits CoVC 10-43-06 exhibited better response under double shade net with a greater number of leaves (3.60) in polyhouse condition.
The response of thirteenth genotype CoVC 10-38-07 under different hardening conditions. The single shade net showed better response for chlorophyll content, root volume, number of roots and root length except shoot length (35.67), number of leaves (3.13) and mortality ( 2 1. 6 3 %) and for which double shade net is the best.
The fourteenth genotype Co 86032 under different hardening conditions. Among three hardening conditions single shade net showed better response for the genotype Co 86032 in terms of chlorophyll content, root volume, number of roots and root length and exhibited higher values for shoot length (21.13) and number of leaves (3.60) under double shade net however, polyhouse is found to be the best for mortality percentage and found that higher temperature (>38 0 C) is not optimal for chlorophyll biosynthesis as it is partly due to impairment of 5- aminolevulinic acid biosynthesis ( 7 8 %) and Protochlorophyllide.
The fifteenth genotype VCF 0517 under different hardening conditions. The double shade net showed better response for characteristics shoot length, number of leaves, chlorophyll content and mortality percentage except root volume (2.00), number of roots (12.37) and root length (7.33), for which single shade net is found best.
The sixteenth genotype CoVC 07-34-05 under different hardening conditions. The Single shade net is optimum for the genotype CoVC 07-34-05 with high root volume (2.40), number of roots (12.23) and root length (6.03). Double shade net showed better response for number of leaves (3.00), chlorophyll content (22.27) and mortality percentage ( 2 1. 9 2 %) and shoot length (24.47) is highest in polyhouse.
To further clarify advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof, which is illustrated in the appended drawings. It is appreciated that these drawings depict only typical embodiments of the invention and are therefore not to be considered limiting of its scope. The invention will be described and explained with additional specificity and detail with the accompanying drawings.
These and other features, aspects, and advantages of the present invention will become better understood when the following detailed description is read with reference to the accompanying drawings in which like characters represent like parts throughout the drawings, wherein:
Figure 1 illustrates a block diagram of all the components used in an apparatus for propagation of sugarcane plant,
Figure 2 illustrates a flow diagram of a method used in the system for propagation of sugarcane, and
Figure 3a-3c illustrates tables of the response of 16 sugarcane genotype under different hardening cage conditions.
Figure 4a-4p show exemplary illustrations of the response of 16 sugarcane genotype under different hardening cage conditions.
Further, skilled artisans will appreciate that elements in the drawings are illustrated for simplicity and may not have been necessarily been drawn to scale. For example, the flow charts illustrate the method in terms of the most prominent steps involved to help to improve understanding of aspects of the present invention. Furthermore, in terms of the construction of the device, one or more components of the device may have been represented in the drawings by conventional symbols, and the drawings may show only those specific details that are pertinent to understanding the embodiments of the present invention so as not to obscure the drawings with details that will be readily apparent to those of ordinary skill in the art having benefit of the description herein.
For the purpose of promoting an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the drawings and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the illustrated system, and such further applications of the principles of the invention as illustrated therein being contemplated as would normally occur to one skilled in the art to which the invention relates.
It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the invention and are not intended to be restrictive thereof.
Reference throughout this specification to "an aspect", "another aspect" or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrase "in an embodiment", "in another embodiment" and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.
The terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process or method that comprises a list of steps does not include only those steps but may include other steps not expressly listed or inherent to such process or method. Similarly, one or more devices or sub-systems or elements or structures or components proceeded by "comprises...a" does not, without more constraints, preclude the existence of other devices or other sub-systems or other elements or other structures or other components or additional devices or additional sub-systems or additional elements or additional structures or additional components.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The system, methods, and examples provided herein are illustrative only and not intended to be limiting.
Embodiments of the present invention will be described below in detail with reference to the accompanying drawings.
Figure 1 illustrates a block diagram of all the components used in an apparatus for propagation of sugarcane plant. The apparatus comprises of a basal medium 100 in a sterilized glassware, a container 102, a portray 104, a hardening cage 106, a thermometer 108, a hygrometer 110 and a lux-meter 112.
The basal medium 100 for regenerating roots of sugarcane, wherein the basal medium 100 is provided in the sterilized glassware.
The Basal medium 100 contain amino acids, glucose, and ions (calcium, magnesium, potassium, sodium, and phosphate) essential for cell survival and growth. I-glutamine is an amino acid that is essential for protein and nucleic acid synthesis and energy production in a cell culture.
The sterilized glassware containing a defined quantity of double distilled water and a plurality of chemicals and stock solutions in required amount.
Sterilization refers to any process that removes, kills, or deactivates all forms of life (in particular referring to microorganisms such as fungi, bacteria, spores, unicellular eukaryotic organisms such as Plasmodium, etc.) and other biological agents like prions present in a specific surface, object or fluid.
The glassware is preferably a bowl, a tumbler, a plate or a glass.
The container 102 is autoclaved at 110-130 degree Celsius for 15-25 minutes and stored under light for about 15-20 hours and under dark for about 7-9 hours for inducing shoot and forming roots.
The term 'autoclave' herein refers to a machine that provides a physical method of sterilization by killing bacteria, viruses, and even spores present in the material put inside of the vessel using steam under pressure. Autoclave sterilizes the materials by heating them up to a particular temperature for a specific period of time.
The temperature and the duration of autoclaving differs according to the requirement of the apparatus.
The container 102 includes a pre-sterilized culture bottle holding a liquid medium of 40-50ml, wherein the container 102 is autoclaved at 121 degree Celsius at 15|bpsi for about 15-25 minutes.
The containers 102 are placed under the light source at a height difference of 25-30cm, wherein subculturing is done after every two three weeks.
The container 102 is accommodated with well rooted with at least 16 genotypes, wherein the genotypes are obtained through an in-vitro culture of apical meristem from top shoot of each of the plurality of sugarcane stock plants.
The protray 104 is for accommodating at least a heterotrophic micro propagated plant containing a mixture of soil, sand and cocopeat in a defined ratio, wherein the ratio of soil, sand and cocopeat in the portray is 1:1:1 respectively.
The hardening cage 106 is used for culturing the heterotrophic micro propagated plants into an autotrophic and acclimatized sturdy plant, wherein the hardening cage 106 comprises of a shade net for allowing light to reach the plants for growth.
The term hardening is a process where heterotrophic in vitro cultured delicate plantlets become autotrophic and acclimatized sturdy plants. This hardening process is carried out in the structures called hardening cage 106.
The hardening cage 106 may be a single shade cage, a double shade cage or a polyhouse. The hardening cage 106 used herein is a single shade cage. The shade net house is constructed with a green shade net permitting 50 per cent light and the average temperature, relative humidity and light intensity were 28.3 0C, 63.20 per cent and 7522 lux, respectively. The thermometer 108, the hygrometer 110 and the lux meter 112 are used to monitor temperature, relative humidity and light intensity respectively in all three hardening cage 106 structures.
Figure 2 illustrates a flow diagram of a method used in the system for propagation of sugarcane.
The method for sugarcane propagation, the method comprises of:
Step 202 discloses about regenerating roots of sugarcane in a basal medium 100, wherein the steps of regeneration comprising of:
Step 202a discloses about adding a defined quantity of double distilled water, a plurality of chemicals and stock solutions in required amount into a sterilized glassware.
Step 202b discloses about autoclaving at least a container 102 at 110 130 degree Celsius for 15-25 minutes and storing under light for about -20 hours and under dark for about 7-9 hours for inducing shoot and forming roots.
Step 202c discloses about accommodating at least a heterotrophic micro-propagated plant in at least a portray, wherein the protray 104 contains a mixture of soil, sand and cocopeat in a defined ratio.
Step 204 discloses about culturing the heterotrophic micro-propagated plants into an autotrophic and acclimatized sturdy plant inside a hardening cage 106, wherein the hardening cage 106 comprises of a shade net for allowing light to reach the plants for growth.
Figure 3a-3c illustrates a tabular representation of the response of 16 sugarcane genotype under different hardening cage 106 conditions for observing the growth characteristics.
The different hardening conditions are single shade net, double shade net and polyhouse.
The shade net house is constructed with green shade net permitting per cent light and the average temperature, relative humidity and light intensity were 28.3 0 C, 63.20 per cent and 7522 lux, respectively.
The double shade net hardening cage 106 structure made with two layers of green shade net and the average temperature, relative humidity and light intensity were 31.39 0C, 68.88 per cent and 5210.3 lux, respectively.
The Polyhouse hardening structure constructed with UV stabilized poly sheet and the average temperature, relative humidity and light intensity were 38.03 0C, 78.14 per cent and 31685.7 lux, respectively.
The thermometer 108, hygrometer 110 and lux meter 112 are used to monitor temperature, relative humidity and light intensity respectively in all three hardening cage 106 structures.
The characteristics considered for monitoring the growth are shoot length, number of leaves, chlorophyll content (mg/g) and root characteristics: number of roots, root length, root volume and survivability.
Figure 3a shows the effect of hardening environment on Mortality (%) and Chlorophyll content (mg/g) of in vitro raised sugarcane plants.
Figure 3b shows the effect of hardening environment on Root volume (cm3) and Root length (cm) of in vitro raised sugarcane plants.
Figure 3c shows the effect of hardening environment on shoot length (cm) and no. of leaves of in vitro raised sugarcane plants. Figure 4a 4p show exemplary illustrations of the response of 16 sugarcane genotype under different hardening cage conditions.
The response of a first genotype C85004 under different hardening cage 106 conditions. For the first genotype Co 85004 single shade net showed better response for all the characteristics except chlorophyll content which recorded highest value (21.80 mg/g) in double shade net and better survivability ( 2 1.5 0 %) in polyhouse.
The response of second genotype Co0323 under different hardening cage 106 conditions. Single shade net is also found optimum for hardening of Co 0323 considering all characteristics measured except mortality percentage (7.5 0 %) for which double shade net was the best.
The response of third genotype CoVC 07-06-05, fourth genotype CoVC 09-65-04, a fifth genotype CoVC 99463, a sixth genotype CoVc 09 61-07, and a seventh genotype CoVC 00964 under different hardening cage 106 conditions. single shade net was found optimal for all shoot morphological characteristics.
The response of eighth genotype Co 99004 under different hardening cage 106 conditions. The optimum response for hardening of Co 99004 is in single shade for all characters except shoot length (36.93) and number of leaves (3.33) for which polyhouse was found to be the best. The degree of adaptation is to be increased by growing shoots in reduced humidity before transfer, but shoot growth and rooting capacity may decline progressively as humidity is lowered.
The response of ninth genotype Co62175 under different hardening cage 106 conditions. The single shade net showed better response for all characteristics except for the number of leaves (Table 5). For number of leaves the highest value is observed (3.07) in double shade net and better survivability recorded ( 3 2 .5 2 %) in polyhouse in the genotype Co 62175.
The response of tenth genotype CoVC 10-65-01 under different hardening conditions. Under single shade net, CoVC 10-65-01 showed better response for all characteristics except shoot length and for which highest (39.60) value is observed in double shade net with better survivability (7. 3 5%) recorded in polyhouse.
The response of eleventh genotype CoT 12368 under different hardening conditions. The genotype CoT 12368 showed better response under single shade net for the four characteristics chlorophyll content, root volume, number of roots and root length. For shoot length (40.27), number of leaves (3.13) and mortality percentage (10.1 7% ) polyhouse was found to be best.
The response of twelfth genotype CoVC 10-43-06 under different hardening conditions. Single shade net is found better for the genotype CoVC 10-43-06 considering chlorophyll content, root volume, number of roots and root length except shoot length (27.40) and mortality percentage (17. 48 %). For these two traits CoVC 10-43-06 exhibited better response under double shade net with a greater number of leaves (3.60) in polyhouse condition.
The response of thirteenth genotype CoVC 10-38-07 under different hardening conditions. The single shade net showed better response for chlorophyll content, root volume, number of roots and root length except shoot length (35.67), number of leaves (3.13) and mortality ( 2 1. 6 3 %) and for which double shade net is the best.
The fourteenth genotype Co 86032 under different hardening conditions. Among three hardening conditions single shade net showed better response for the genotype Co 86032 in terms of chlorophyll content, root volume, number of roots and root length and exhibited higher values for shoot length (21.13) and number of leaves (3.60) under double shade net however, polyhouse is found to be the best for mortality percentage and found that higher temperature (>38 0 C) is not optimal for chlorophyll biosynthesis as it is partly due to impairment of 5- aminolevulinic acid biosynthesis ( 7 8 %) and Protochlorophyllide.
The fifteenth genotype VCF 0517 under different hardening conditions. The double shade net showed better response for characteristics shoot length, number of leaves, chlorophyll content and mortality percentage except root volume (2.00), number of roots (12.37) and root length (7.33), for which single shade net is found best.
The sixteenth genotype CoVC 07-34-05 under different hardening conditions. The Single shade net is optimum for the genotype CoVC 07-34-05 with high root volume (2.40), number of roots (12.23) and root length (6.03). Double shade net showed better response for number of leaves (3.00), chlorophyll content (22.27) and mortality percentage ( 2 1. 9 2 %) and shoot length (24.47) is highest in polyhouse.
The drawings and the forgoing description give examples of embodiments. Those skilled in the art will appreciate that one or more of the described elements may well be combined into a single functional element. Alternatively, certain elements may be split into multiple functional elements. Elements from one embodiment may be added to another embodiment. For example, orders of processes described herein may be changed and are not limited to the manner described herein. Moreover, the actions of any flow diagram need not be implemented in the order shown; nor do all of the acts necessarily need to be performed. Also, those acts that are not dependent on other acts may be performed in parallel with the other acts. The scope of embodiments is by no means limited by these specific examples. Numerous variations, whether explicitly given in the specification or not, such as differences in structure, dimension, and use of material, are possible. The scope of embodiments is at least as broad as given by the following claims.
Benefits, other advantages, and solutions to problems have been described above with regard to specific embodiments. However, the benefits, advantages, solutions to problems, and any component(s) that may cause any benefit, advantage, or solution to occur or become more pronounced are not to be construed as a critical, required, or essential feature or component of any or all the claims.
Claims (10)
1. An apparatus for sugarcane propagation, the apparatus comprises of: a basal medium 100 for regenerating roots of sugarcane, wherein the basal medium 100 is provided in a sterilized glassware containing a defined quantity of double distilled water and a plurality of chemicals and stock solutions in required amount;
at least a container 102 autoclaved at 110-130 degree Celsius for 15-25 minutes and stored under light for about 15-20 hours and under dark for about 7-9 hours for inducing shoot and forming roots;
at least a protray 104 for accommodating at least a heterotrophic micro-propagated plant containing a mixture of soil, sand and cocopeat in a defined ratio; and
a hardening cage 106 for culturing the heterotrophic micro propagated plants into an autotrophic and acclimatized sturdy plant, wherein the hardening cage 106 comprises of a shade net for allowing light to reach the plants for growth.
2. The apparatus as claimed in claim 1, wherein a pH value of the basal medium 100 is maintained using 0.1 NHCI or 0.1N NaOH.
3. The apparatus as claimed in claim 1, wherein the container 102 includes pre-sterilized culture bottle holding a liquid medium of 40 50ml, wherein the container 102 is autoclaved at 121 degree Celsius at 15|bpsi for about 15-25 minutes.
4. The apparatus as claimed in claim 1, wherein the container 102 is stored under light for 16 hours and under dark conditions for 8 hours in a photoperiod condition for shoot induction, shoot multiplication and root formation.
5. The apparatus as claimed in claim 1, wherein the containers 102 are placed under the light source at a height difference of 25-30cm, wherein subculturing is done after every two-three weeks.
6. The apparatus as claimed in claim 1, wherein the container 102 is accommodated with well rooted with at least 16 genotypes, wherein the genotypes are obtained through an in-vitro culture of apical meristem from top shoot of each of the plurality of sugarcane stock plants.
7. The apparatus as claimed in claim 1, wherein the ratio of soil, sand and cocopeat in the portray is 1:1:1, respectively.
8. The apparatus as claimed in claim 1, wherein the shade net house is a single shade net house constructed with green shade net permitting about 50 percent light with average temperature, relative humidity and light intensity of about 28.3 degree Celsius, 63.20 percent and 7522 lux, respectively.
9. The apparatus as claimed in claim 1, wherein a thermometer 108, hygrometer 110 and lux-meter 112 are used to monitor temperature, relative humidity and light intensity, respectively.
10. A method for sugarcane propagation, the method comprises of: regenerating roots of sugarcane in a basal medium 100, wherein the steps of regeneration comprising of: adding a defined quantity of double distilled water, a plurality of chemicals and stock solutions in required amount into a sterilized glassware; autoclaving at least a container 102 at 110-130 degree Celsius for 15-25 minutes and storing under light for about 15-20 hours and under dark for about 7-9 hours for inducing shoot and forming roots; accommodating at least a heterotrophic micro-propagated plant in at least a portray, wherein the protray 104 contains a mixture of soil, sand and cocopeat in a defined ratio; and culturing the heterotrophic micro-propagated plants into an autotrophic and acclimatized sturdy plant inside a hardening cage 106, wherein the hardening cage 106 comprises of a shade net for allowing light to reach the plants for growth.
FIGURE 2 FIGURE 1
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FIGURE 4h 2021102640
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