Anirban Guha

Trees provide shade, but have you ever considered trees themselves needing shade? Our lab is seeking to answer this question. Anirban Guha, who is leading this effort in our lab, was able to sit down with me and answer some questions about the experiment. He joined the lab in April 2019 as a post-doctoral scholar. The experiment is attempting to determine how the trees respond to different light conditions over a period of two to three years with the use of shade nets to manipulate the environmental conditions. The lab records daily, weekly, and monthly results, and will record the yearly results when the time comes. We already know that citrus responds better in partial shade conditions, which improve yield and yield quality and photosynthesis and water status. We think that full sun has an especially bad effect on HLB trees. Anirban explained that the infected plants often cannot take in the full force of Florida sunlight; it provides them with more energy than they have the capacity to process. HLB also stunts root growth, which becomes even more of a problem when high light conditions demand more water and nitrogen than can be taken up by the roots.

The lab is testing whether shading the trees allows them to conserve more energy and require less water and nitrogen, which would help balance their functioning with the disease. Ultimately, the goal is to “[develop an] agricultural system in a way that could modify the environmental cues, and that can lead to better fitness of the plant to help sustain yield and maintain better physical performance.”

The main recipients of this experiment are scientists and citrus growers; Anirban thinks these two groups believe they have different reasons for caring about the results, but he believes their goals are actually similar and the knowledge they seek is complementary. Whether results are sought for economic reasons or a research quest, the ultimate goal is to see the trees become healthier and create more fruit yield—something both the scientific and agricultural communities can agree upon. 

Anirban takes this collaborative approach in his work life as well. The physiology lab collaborates with other CREC labs to study and test infected trees. Their results often work together to create healthier trees. For example, the entomology lab provides information on how insects spread HLB. He desires for more scientists of different disciplines to work together to achieve “functional collaborative research,” which can help the scientific community locally and worldwide. Along with scientists working together to achieve more, he also wants his research to be holistic. He wanted to study trees not just at a cellular level, but “from leaf to whole plant.” After completing his PhD in India, he found the majority of opportunities available there were for study at the cellular level. Anirban was interested in more variety and didn’t want to do what everyone else was doing. He also saw this gap in research as something he could potentially fill back home one day. He believes the study of the whole tree is important because problems tend to be linked to one another and can be better understood when a whole plant approach is taken. He enjoys his work but told me with a good-natured smile that he is not at all attached to the state of Florida and would like to return to India one day. 

Do sprayed antibiotics move inside citrus plants?

There has been discussion lately about how effective are antibiotics in citrus groves. In order for these to take down Las, the bacterium that causes greening, they need to be moved in the vascular system, particularly the phloem, where Las hangs out.  So, are leaf-sprayed antibiotics moved systemically?

This is the question some colleagues and I asked in a recent paper.  We were following up on some of Nian Wang’s research that showed how trunk-injected oxy-tetracycline (which we’ll affectionately call “oxytet”) moved around the citrus plant, eventually reaching all parts.  We had a few questions:

  1. When oxytetracycline is sprayed on the leaves, does it move to other parts of the plant? This question addressed whether there is systemic movement at all.
  2. How much moves? If there is systemic movement, what proportion of what is sprayed is moved.
  3. Does it matter whether you spray on old or young leaves? Some growers try to spray when there is new flush?  New flush has thinner cuticle (the waxy layer on the outside of the leaf). We thought that applying to new flush would allow more oxytet to get into the leaf.

In the same study we looked at some of the effects of heat treatment, but today we’ll just look at oxytet delivery. These trees did not have HLB, this is because we wanted to look at oxytet movement, not efficacy against Las.

What did we find?

  1. Oxytetracycline did move into leaves that did not receive the spray.
  2. How much moved? In one trial we found between 0.36-0.63 of the concentration in the leaves that didn’t receive the application relative to the leaves that were sprayed. But in the next trial we found between 0.27 and 0.34. So, although we did find oxytet moved into leaves that weren’t sprayed, a portion of what was sprayed stayed in the original leaf. The portion that didn’t become systemic probably is useless against Las because it isn’t moving through the vascular system.
  3. It doesn’t matter whether you spray old or young leaves. We found the same concentrations in the leaves regardless of which leaves were sprayed. At first we thought this contradicted some previous work that showed that there was a decrease in delivery of nitrogen as leaves aged. However, taking a closer look at that older work, it turns out that delivery decreases from when leaves emerge until about 6 weeks. However, after this point delivery begins to increase again. This is because, although there is more wax on the outside of the leaf, this wax ages and forms cracks, which probably allow more of whatever is sprayed in.

How did we find this?

Some plants we removed all new flush, some we removed all old flush, and some we left all the leaves. We covered about 1/4 of the canopy of small trees with impermeable plastic. Then we sprayed the rest of the canopy. After the spray had dried, we removed the plastic. About 3 weeks later we sampled both the leaves that were directly sprayed and those that weren’t. Then we tested each for oxytetracycline content.

Is it enough?

We still don’t know whether the concentrations that made it do the unsprayed leaves were enough to reduce the Las levels, because we actually don’t know how much oxytet it takes to bring Las down in the plant. This study didn’t address streptomycin, the other antibiotic that is labeled to use against Las, so we don’t know whether it would move similarly.

How do citrus trees ‘decide’ when to flush?

Because the life cycles of two important pests, Asian citrus psyllid and citrus leaf miner, are closely linked to the emergence of new flush.  Growers could benefit from spraying only when there is new flush.  But if the late flush is spread out over a month or even more, then what’s a body to do?

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Asian citrus psyllid nymphs feeding on young flush, not old leaves.

There may be ways we can manipulate the timing of flush directly.  But to do that we need to understand how the plant regulates its own growth patterns.

Citrus trees have a different growth cycle than most common fruits.  Like many fruits, they flower in the spring and then fruit develops over the next several months.  Deciduous plants add vegetative growth (leaves and shoots) during a solid, extended period of the year.  Citrus plants grow in several cycles of vegetative flushes over the year.  The number and the size of these flushes depends on the environment.  In Florida there are 2-3 vegetative flushes per year.  Usually this begins with a flush in May, with later flushes in July and September or October.  These flushes become less coordinated from plant to plant in a grove as the year waxes on, with May flush being relatively concentrated and the late flush being very sporadic.

Citrus plants’ “distribution of wealth”

The plant has to balance the needs of the leaves for water and nutrients with the needs of fruits and roots for sugars.  This gets even more complicated when the plant has to send sugars to a new shoot to new flushes that will need to grow more than 1 cm in a day.  So the plant needs to to start flushing at just the right time so that its fruits and roots don’t starve.  It also needs to not start a new flush when the old one is not yet mature.  What researchers in the 1990s observed is that root growth stops when shoots are growing.

How do they do it?

We humans have the benefit of brains that coordinate all the different ends of our body, so that our heart beast the right amount for our feet to be able to run.  Plants don’t have benefit of brains, so they rely on chemical signals.  These signals can be molecules that move from roots to leaves or that accumulate in one part of the plant when the other part stops using that compound.  Most of these signals are called phytohormones, because their effects are very large compared their very small concentrations.  Growing shoots send out signals called auxins that keep other buds from growing so that there aren’t too many flushes.  Meanwhile, gibberelic acid and cytokinins coordinate between roots and shoots. Not all of these dynamics have been completely explained in the case of citrus flush phenology, but we are gaining ground!

Why does it matter?

Knowing the “code” the roots, leaves, and shoots use to communicate, allows us to grab the microphone and give a few orders. Thus it may be possible to manage the flush timing, to decrease its sporadic nature and improve the efficacy of our pest management, ultimately getting healthier plants with greater yields.