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Figure 2. Romaine lettuce plants in a field<br />

showing variation in severity typical of lettuce<br />

dieback including stunted growth, as well as<br />

yellowing and necrosis of outer leaves.<br />

Since the late 1990s, lettuce dieback has<br />

been believed to be caused by infection of<br />

lettuce plants with either of two viruses<br />

from the genus Tombusvirus; tomato<br />

bushy stunt virus (TBSV) and Moroccan<br />

pepper virus (MPV). These viruses are<br />

absent from healthy lettuce but have<br />

been found regularly in association with<br />

lettuce dieback disease. However, there<br />

have been numerous situations in which<br />

neither virus was found in association<br />

with obvious disease symptoms. Furthermore,<br />

it has not been possible to<br />

consistently and easily reproduce disease<br />

symptoms when lettuce is inoculated<br />

with either virus in a laboratory setting,<br />

raising the possibility that an additional<br />

virus may contribute to causing lettuce<br />

dieback disease.<br />

In an attempt to identify a possible<br />

additional virus contributing to lettuce<br />

dieback disease, high throughput<br />

sequencing (HTS) was used on several<br />

lettuce plants exhibiting dieback symptoms,<br />

which led to the identification of<br />

a new virus consistently associated with<br />

diseased plants but not with healthy<br />

lettuce plants. This novel virus was most<br />

closely related to a recently identified<br />

and poorly characterized virus from watermelon<br />

in China, watermelon crinkle<br />

leaf associated virus, which was found<br />

using the same HTS approach.<br />

The newly identified lettuce virus, tentatively<br />

named lettuce dieback associated<br />

virus (LDaV) shares an extremely low<br />

genetic relationship with the watermelon<br />

virus, which suggests that although the<br />

two viruses are related, they are very<br />

distantly related to one another. Using<br />

a combination of HTS and traditional<br />

DNA sequencing the genome of the<br />

new virus, LDaV, was assembled and<br />

Figure 3. Romaine lettuce leaf from the inner<br />

portion of a head showing star-shaped chlorotic<br />

flecking in veins characteristic of lettuce dieback<br />

disease on romaine.<br />

methods were developed to allow rapid<br />

detection of the virus from lettuce leaf<br />

extracts using RT-PCR, a routine laboratory<br />

diagnostic method. LDaV has now<br />

been found not only in lettuce showing<br />

dieback symptoms collected recently, but<br />

it has also been found in older archived<br />

samples of lettuce nucleic acid collected<br />

from plants showing dieback symptoms<br />

over the past 20 years, including many<br />

that also contained MPV or TBSV. To<br />

date, LDaV has not been found in healthy<br />

lettuce plants. Interestingly, genetic<br />

comparison showed that LDaV isolates<br />

collected from coastal California production<br />

regions are closely related to one<br />

another, and desert isolates from Arizona<br />

and Imperial Valley, California also are<br />

closely related to one another. However,<br />

coastal and desert isolates differ genetically<br />

from one another, suggesting<br />

perhaps some regional adaptation of the<br />

virus to plants grown under the different<br />

climatic conditions.<br />

Further research will clarify the role of<br />

LDaV in lettuce dieback disease and how<br />

it relates to the two tombusviruses, MPV<br />

and TBSV, that have long been linked<br />

to the disease. Studies to date, however,<br />

strongly suggest a role for LDaV in<br />

lettuce dieback disease development, and<br />

research is in progress to clarify the ability<br />

of LDaV to produce lettuce dieback<br />

symptoms when inoculated to lettuce<br />

plants, as well as whether or not the new<br />

virus can infect lettuce plants carrying a<br />

gene for resistance to lettuce dieback.<br />

Comments about this article? We want<br />

to hear from you. Feel free to email us at<br />

article@jcsmarketinginc.com<br />

<strong>July</strong> / <strong>August</strong> <strong>2020</strong> www.progressivecrop.com 31

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