What is acclimatization in plant

tissue culture?
 The greenhouses or fields have relatively lower relative humidity, higher light level,
and an environment full of pests and microorganisms around. These conditions pose
extreme stress to plants cultured under in vitro environments.
 Therefore, a successful tissue culture protocol for a plant species also involves the
efficient transfer of plantlets from culture vessels to the ex vitro (field/greenhouse)
conditions.
 In the tissue culture process, you also need to care about the mother plants
which you use for obtaining explants.
 As much as acclimatization is important for a successful result, taking
precautions to prevent contaminations at the beginning of the process is also
crucial.
What is acclimatization?
Acclimatization simply means the adaptation of plantlets to a new environment.

 Plantlets or shoots in the culture vessels are accustomed to a different micro-

environment.
 You customize this micro-environment in a way that the developing plants
experience minimal stress and optimum conditions to grow and multiply.
 When you transfer these plantlets from the laboratory conditions to the soil, you
expose them to different abiotic and biotic stresses.
 Hence, you need to develop a stepwise acclimatization protocol for your plants
to adapt to the natural environment.
Why do you need a stepwise process for acclimatization?
In the culture vessels, your plantlets are growing in controlled conditions,
such as:
•Low level of light;
•High level of humidity;
•Aseptic conditions; and
•Nutrient medium with a substantial amount of sugar.
 Plants growing in such conditions cannot survive the environmental conditions
when they are directly transferred in the open.
 Because of this reason, the stage of acclimatization is a major bottleneck in the
micropropagation of many plant species and requires a carefully curated
stepwise process to adapt to an open environment.
What are the problems faced by in vitro plants during acclimatization?

 The presence of high humidity in the culture vessels leads to abnormal functioning of
stomata upon transfer to soil. This directly influences the rate of transpiration in
plants or in other words, it affects the rate at which a plant absorbs and utilizes water
for growth.
 Due to low light intensity, low carbon dioxide concentration, and high sugar in the
media, plantlets are accustomed to a relatively low rate of photosynthesis, which
further leads to slow growth.
 As plantlets grow in sterile media, they possess weaker roots when compared with
field plants. These weak roots experience difficulty in uptaking nutrients from the soil.
 When plantlets grow in excess amount of plant growth regulators, they show
abnormalities in morphology upon transfer to greenhouse conditions.
 When plantlets are transferred from low light intensity and temperature to broad
spectrum sunlight and higher fluctuating temperature, then they experience
charring of leaves (burning of leaves/browning of plant tissues) and wilting of
plants.
 Direct transfer of plantlets to sunlight can also inhibit the process of
photosynthesis which is necessary for energy production.
 Another major cause of mortality in tissue cultured plantlets is their sudden
exposure to microflora present in the soil. As these plants are not sufficiently
resistant to such organisms, they get infected easily and do not survive.
Hence, plants cultivated in vitro are different from field-grown plants
and require more attention while adapting to a new environment.
How can you acclimatize your plants?
Let's discuss the measures you can take to acclimatize your plantlets. Also, what factors
do you need to keep in mind to ensure the survival of plantlets?
 First, it is necessary to transfer the plants from a tissue culture environment to a
greenhouse providing the same environment in which it was cultured in the lab.
 You need to transfer the plantlets to a potting mix, irrigated with inorganic nutrient
solution. A variety of potting mixes are available nowadays including peat, vermiculite,
soil, sand, etc.
 You can also keep the culture vessels with loose lids for a few days in the
greenhouse.
 You can leave the plantlets in shade for 3-6 days under diffused natural light. This
would help them to establish under new environment.
 According to scientists, acclimatization should occur in 2-3 phases where you
gradually expose plantlets first to greenhouse and then to field conditions.
 In order to adapt plants from high to low humidity, you need to keep them in shade
with loose plugs for a week or two. Then you should transfer them to pots containing
sterile soil and sand mixture. You can cover these pots with polybags.
 You can also precondition your rooted plantlets in different sucrose solutions (20-
30g/L of concentrations) before transferring to a potting mixture. This would increase
the growth of shoots.
 In several studies, it is also suggested to add anti-transpirants like
'paclobutrazol' in the rooting medium. This helps plantlets to have normal
functioning of stomata, thickened roots and also reduces wilting.
 You need to customize these steps for each plant variety based on the
plant's growth requirements.
 The steps mentioned in this section will shock your plantlets at first as the
growth media will change from inorganic media to organic soil. But, this will
also activate the photosynthetic activity of the plantlets and prepare them to
withstand the natural environment.
 By keeping these steps in mind, you can ensure a good start for your
plantlets in field conditions, be it for a small or a large-scale production.