(if you’re here for the first time look at Blogs 1 -11 in earlier posts )
Maddie went back to her lab work after the seminar. It took her three weeks of running gels practically every night in order to complete the process. On a gel, a column of stained protein bands represented each of the wild beans. She had numbered every protein band on the gel from top to bottom just as one would number the lines on a piece of paper. Some lines showed blank spaces and other lines were filled by a stained protein band. The data for each of the wild varieties had been entered in the computer as she got the gels done; each had a unique set of proteins. The question was, did the resistant ones all have a protein band in common?
“Tonight’s the night! Okay baby, now let’s see what you got,” she said to the empty computer room. The blue screen lit up and Maddie entered the protein data into the computer. Next, she pulled up the file with the corresponding disease ratings for bacterial blight obtained over the summer, and began setting up the statistical analysis.
“Please, give me a significant difference,” she implored, crossing her fingers as the computer crunched the numbers. It was getting late and she wanted to have something to report at the research group meeting tomorrow. She wanted to report some results! Preferably, that lines that contained a certain protein band had significantly higher resistance than the ones that didn’t.
When she heard the hum of the printer she jumped up, eager to scan the results one at a time, careful not to take her eyes off the printout. She took a deep breath, bracing herself. The numbers came out drop by drop. At the end her hopes were dashed. There was no clear-cut relationship between any of the 34 protein bands and resistance to bacterial blight.
She tore the sheet off the digital printer and went over it again.
“Nope, you can’t say there’s a significant difference there. The means are the same whether the protein band is present or absent.” She sighed. “Whatever the mechanism of resistance is, it’s definitely not in these proteins.”
She started running the analysis for anthracnose next; mumbling to herself, “Well, you still got four chances left to hit the jackpot, so don’t despair, baby.”
But after finding no significant difference in the case of anthracnose or BCMV either, she began to get worried. She ran her fingers through her hair. “All that work collecting those disease ratings! Multiple readings!” she wailed, fighting a growing sense of discouragement. It was going on twelve o’clock; at this rate she was averaging an hour per disease.
“Oh please, please give me something,” she murmured as she set up the Bean Lesion Virus analysis. Bean Lesion Virus was a disease that showed up as lesions on the exterior of the pods. It occurred close to maturity and so there was a good possibility that a seed compound could be closely associated with resistance because it might provide toxicity to the virus.
“Come-on, you’re my best hope. If you don’t have it, then I’m sunk. I’ll have to start this thesis all over again…” She scanned the printout as it was coming off the laser printer. When the analysis confirmed that there was a highly significant difference for band 23 she stood motionless, hardly able to believe her eyes. There was no room for doubt.
“Yes!” she yelled, and began hopping around the deserted computer room. It didn’t matter that it was one o’clock in the morning.
With significance this high, there was definitely something to investigate. Perhaps there was even a direct relationship. Perhaps band 23 was the actual resistance compound itself, and not just a marker!
She went on to set up the analysis for Rusty Leaf Spot, still hoping for something, but a little less desperate than before. As the printout began emerging she kept her eye on each band. Band 1 had no significant difference, band 2 had nothing either, neither did band 3.
Every subsequent band went by with similar numbers. “I don’t believe it,” she wailed, and then glanced down to see the last band, band 34, showing a highly significant difference!
“Oh my God! Thank you, thank you!” This time she didn’t dance around, she just sat there and rubbed her eyes, as much from emotion as from lack of sleep. She’d better get home for a few hours rest. She wanted to be at her best for the research group meeting at noon. The taste of success was sweet in her mouth.
The Bean Group met in the teaching lab on the first floor every Friday. Maddie walked in on the group spread around the black-topped lab bench. Most everyone had brought their lunch. Dr. Gates currently had ten graduate students, all in varying stages of their Masters or Ph.D. degrees. All were males except for Maddie and Emily Mbasa.
Maddie waited until Dr. Gates or Edgar had covered most other items of business and reports.
“Excuse me.” She cleared her throat, feeling the heat rise up in her face. She became so self-conscious when speaking up at these meetings.
“Yes, Maddie. Do you have something?” Dr. Gates acknowledged her.
She cleared her throat, and then began, “I wanted to report that I ran an analysis yesterday on the five disease ratings and the 34 seed protein bands.” She paused for a deep breath. “I found no significant differences for bacterial blight, anthracnose, and BCMV. I did, however, find a significant relationship between band 23 and Bean Lesion Virus and also for band 34 and Rusty Leaf Spot.”
There were little murmurs of approval around the table and then a barrage of questions.
“How significant were the differences between means?” Dr. Gates wanted to know.
Maddie moved to the blackboard and wrote out the means. “Very highly significant, as you can see. This is Bean Lesion Virus and this is for Rusty Leaf Spot.”
“Wow,” someone piped in.
Edgar asked, “What about white mold and powdery mildew? I thought you were going to rate for those too. The Bean Growers Association is really looking for resistance to those two.”
Maddie didn’t want a pall cast over her good news, “Well, I didn’t really have a good infestation in my plots. In fact I don’t know if anybody did this year. Did you?”
“No, not really.” Edgar had to admit.
“Maddie, are those bands identified yet?” Alex Vieira asked.
“No. They’re not among any of the proteins we’ve worked on in the past.”
“Are you sure?” Paul Zwiteck challenged.
“Yeah, I looked!” Maddie asserted. “They don’t exist in the profile of the cultivated lines I used for controls[i] either. But I have to—”
“What about the literature? Did you search the literature to see if anybody has classified them?” Paul interrupted again; the excitement about the possibility of a completely new discovery was mounting.
Maddie backed down at this point saying, “No, I was just going to say that I have to do that still. I just came up with these results last night so I didn’t have time—”
“What’s your next step?” Rusty Cameron asked.
“Well, I start crosses next week. I’ll cross all the lines that have these bands with the cultivated lines—”
“You should look for a wild line with both bands, that way you’ll save having to do another cross to combine the two later,” Rusty interjected again.
“Uh-hum.” It wasn’t such a bad suggestion but it was premature. So little was known about the lines with proteins 23 and 34. She didn’t want to end up crossing the wrong ones or exclude any from the crosses at this point.
Maddie thought she saw Emily Mbasa raise her hand tentatively but then retreat into silence. Emily never spoke at these meeting unless directly addressed. And then she did so very quietly. Maddie wondered sometimes why she didn’t try harder to assert herself. She felt that to the rest of the group Emily was not even viewed as a player.
Later that day Emily came over to her desk and placed a paper in her hands saying, “Perhaps you will find this useful. It is a technique for extracting proteins out of vegetative material.”
“Emily, this is great! That’s exactly what I thought of doing, trying to see if this protein band was also present in the pod tissue.”
“Yes, because you see, it may have a different form than the seed protein. The one in the seed may be inert, while the one in the green pod may be active, so if you’re going to investigate the actual compound that is producing toxicity to the virus, then you should work with the pod protein.”
“Yeah, I agree. I wonder who can help me with the extraction?”
“Why don’t you try talking to Dr. Gillian in the Plant Pathology department?”
“I will. Thank you for the paper, Emily.” The African woman turned to go off to her own cubicle.
“Emily, wait!” Maddie called.
Emily glanced back and waited for Maddie to speak.
“How come…was there some particular reason you didn’t bring this up in the meeting? It seemed to me you were going to.”
Emily hesitated a moment then said, “I do not feel comfortable speaking at these meetings.”
“Because I feel everything I would say would sound stupid. And one would have to shout in order to be heard. I am not good at shouting.”
Maddie smiled. “You know, I’m so glad you said that, because I feel the same way. I get so self-conscious when I open my mouth. And all the guys, they seem to have all the answers, to know it all.”
“Ah! They do not know it all! They only appear to!” Emily burst out, and then covered her mouth in embarrassment. Her outburst was a departure from her sedate and prim demeanor.
“Girl, you got that right!” Something passed between them and they both smiled.
[i] Controls – any variety of bean that is commonly used as a baseline to compare against.
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